U.S. patent number 8,575,176 [Application Number 13/353,018] was granted by the patent office on 2013-11-05 for heteroaromatic pyrazinoylguanidine sodium channel blockers.
This patent grant is currently assigned to Parion Sciences, Inc.. The grantee listed for this patent is Michael Ross Johnson. Invention is credited to Michael Ross Johnson.
United States Patent |
8,575,176 |
Johnson |
November 5, 2013 |
Heteroaromatic pyrazinoylguanidine sodium channel blockers
Abstract
Polyaromatic sodium channel blockers represented by the formula:
##STR00001## are provided where the structural variables are
defined herein. The invention also includes a variety of
compositions, combinations and methods of treatment using these
inventive sodium channel blockers.
Inventors: |
Johnson; Michael Ross (Chapel
Hill, NC) |
Applicant: |
Name |
City |
State |
Country |
Type |
Johnson; Michael Ross |
Chapel Hill |
NC |
US |
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Assignee: |
Parion Sciences, Inc. (Durham,
NC)
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Family
ID: |
40998513 |
Appl.
No.: |
13/353,018 |
Filed: |
January 18, 2012 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20120116083 A1 |
May 10, 2012 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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12393252 |
Feb 26, 2009 |
8124607 |
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61031466 |
Feb 26, 2008 |
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Current U.S.
Class: |
514/255.06;
546/148; 546/152; 544/407 |
Current CPC
Class: |
A61P
1/00 (20180101); C07D 241/26 (20130101); A61P
13/00 (20180101); C07D 405/14 (20130101); A61P
27/02 (20180101); A61P 43/00 (20180101); A61P
11/02 (20180101); C07D 241/32 (20130101); A61P
11/06 (20180101); A61P 11/08 (20180101); C07D
405/12 (20130101); A61P 11/00 (20180101); A61P
9/00 (20180101); A61P 35/00 (20180101) |
Current International
Class: |
A61K
31/4965 (20060101) |
Field of
Search: |
;514/255.06 ;544/407
;546/148,152 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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WO 2007/071396 |
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Jul 2007 |
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WO |
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Other References
Jordan, V. C. Nature Reviews: Drug Discovery, 2, 2003, 205. cited
by examiner .
Dorwald, F. Zaragoza. Side Reactions in Organic Synthesis: A Guide
to Successful Synthesis Design, Weinheim: Wiley-VCH Verlag GmbH
& Co. KGaA, 2005, Preface. cited by examiner .
Chawla, et al. Curr. Res. & Info. Pharm. Sci. (CRIPS), 5, 1,
2004, 9-12. cited by examiner .
Office Action issued Jan. 17, 2012 in European Patent Application
No. 09 747 034.8-2101. cited by applicant .
U.S. Appl. No. 60/495,725, filed Aug. 18, 2003, Johnson. cited by
applicant .
U.S. Appl. No. 60/495,720, filed Aug. 18, 2003, Johnson. cited by
applicant .
U.S. Appl. No. 60/495,712, filed Aug. 18, 2003, Johnson. cited by
applicant .
U.S. Appl. No. 60/602,312, filed Aug. 18, 2004, Johnson. cited by
applicant .
U.S. Appl. No. 60/602,327, filed Aug. 18, 2004, Johnson. cited by
applicant .
U.S. Appl. No. 60/812,091, filed Jun. 9, 2006, Johnson. cited by
applicant .
U.S. Appl. No. 60/812,077, filed Jun. 9, 2006, Johnson et al. cited
by applicant .
U.S. Appl. No. 60/812,078, filed Jun. 9, 2006, Johnson. cited by
applicant .
U.S. Appl. No. 60/842,669, filed Sep. 7, 2006, Johnson et al. cited
by applicant .
U.S. Appl. No. 60/842,963, filed Sep. 8, 2006, Johnson et al. cited
by applicant .
U.S. Appl. No. 60/845,171, filed Sep. 18, 2006, Johnson et al.
cited by applicant .
U.S. Appl. No. 11/696,003, filed Apr. 3, 2007, Johnson. cited by
applicant .
U.S. Appl. No. 60/909,818, filed Apr. 3, 2007, Johnson et al. cited
by applicant .
U.S. Appl. No. 60/909,802, filed Apr. 3, 2007, Johnson et al. cited
by applicant .
U.S. Appl. No. 60/978,887, filed Oct. 10, 2007, Boucher et al.
cited by applicant .
U.S. Appl. No. 60/978,874, filed Oct. 10, 2007, Boucher et al.
cited by applicant .
U.S. Appl. No. 60/987,663, filed Nov. 13, 2007, Johnson et al.
cited by applicant .
U.S. Appl. No. 60/013,387, filed Dec. 13, 2007, Johnson et al.
cited by applicant .
U.S. Appl. No. 61/030,313, filed Feb. 21, 2008, Johnson. cited by
applicant .
U.S. Appl. No. 61/031,466, filed Feb. 26, 2008, Johnson. cited by
applicant .
U.S. Appl. No. 12/171,867, filed Jul. 11, 2008, Johnson et al.
cited by applicant .
U.S. Appl. No. 12/171,897, filed Jul. 11, 2008, Johnson et al.
cited by applicant .
U.S. Appl. No. 61/079,989, filed Jul. 11, 2008, Boucher et al.
cited by applicant .
New Zealand Examination Report issued Apr. 16, 2012 in patent
application No. 586940. cited by applicant .
Office Action in Japanese Application No. 200980106218.2 dated Sep.
27, 2012. cited by applicant .
U.S. Appl. No. 13/467,618, filed May 9, 2012, Johnson et al. cited
by applicant .
U.S. Appl. No. 13/492,711, filed Jun. 8, 2023, Johnson et al. cited
by applicant .
U.S. Appl. No. 13/533,911, filed Jun. 26, 2012, Johnson. cited by
applicant .
U.S. Appl. No. 13/669,822, filed Nov. 6, 2012, Johnson. cited by
applicant .
Japanese Office Action dated Sep. 20, 2011 in corresponding
Japanese Application No. 200980106218.2. cited by applicant .
Chinese Office Action in corresponding Application No.
200980106218.2 dated Jul. 1, 2013. cited by applicant .
European Office Action in corresponding Application No. 09747034.8
dated Aug. 21, 2013. cited by applicant.
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Primary Examiner: Willis; Douglas M
Attorney, Agent or Firm: Oblon, Spivak, McClelland, Maier
& Neustadt, L.L.P.
Parent Case Text
CONTINUING APPLICATION INFORMATION
This application is a Continuation of U.S. patent application Ser.
No. 12/393,252 filed on Feb. 26, 2009, now U.S. Pat. No. 8,124,607
now allowed.
CONTINUING APPLICATION DATA
This application claims priority to U.S. provisional application
Ser. No. 61/031,466 filed on Feb. 26, 2008, and incorporated herein
by reference.
Claims
What is claimed is:
1. A compound represented by the formula III: ##STR00068## and
pharmaceutically acceptable salts thereof, wherein: A.sup.2 is
represented by the formula: ##STR00069## wherein one Q is a
nitrogen atom, five Q are carbon atoms and one Q is C--R.sup.5;
R.sup.5 is --O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00070##
2. The compound of claim 1, which is an acid addition salt of an
inorganic acid or an organic acid selected from the group
consisting of hydrochloric acid, hydrobromic acid, sulfuric acid,
phosphoric acid, nitric acid, acetic acid, oxalic acid, tartaric
acid, succinic acid, maleic acid, fumaric acid, gluconic acid,
citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid,
palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic
acid, methanesulfonic acid, p-toluenesulfonic acid,
naphthalenedisulfonic acid, polygalacturonic acid, malonic acid,
sulfosalicylic acid, glycolic acid, salicylic acid, stearic acid,
phthalic acid, mandelic acid, and lactic acid, and racemates,
enantiomers, diastereomers, and tautomers thereof.
3. A pharmaceutical composition, comprising the compound of claim 1
or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
4. A method of promoting hydration of mucosal surfaces, comprising:
administering an effective amount of the compound of claim 1 or a
pharmaceutically acceptable salt thereof to a mucosal surface of a
subject.
5. A method of blocking sodium channels, comprising: contacting
sodium channels with an effective amount of the compound of claim 1
or a pharmaceutically acceptable salt thereof.
6. A method of inducing sputum, comprising administering to a human
in need of increased mucociliary clearance or mucosal hydration an
effective amount of an osmolyte and the compound of claim 1 or a
pharmaceutically acceptable salt thereof.
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to sodium channel blockers. The
present invention also includes a variety of methods of treatment
using these inventive sodium channel blockers.
2. Description of the Background
The mucosal surfaces at the interface between the environment and
the body have evolved a number of "innate defense", i.e.,
protective mechanisms. A principal form of such innate defense is
to cleanse these surfaces with liquid. Typically, the quantity of
the liquid layer on a mucosal surface reflects the balance between
epithelial liquid secretion, often reflecting anion (Cl.sup.-
and/or HCO.sub.3.sup.-) secretion coupled with water (and a cation
counter-ion), and epithelial liquid absorption, often reflecting
Na.sup.+ absorption, coupled with water and counter anion (Cl.sup.-
and/or HCO.sub.3.sup.-). Many diseases of mucosal surfaces are
caused by too little protective liquid on those mucosal surfaces
created by an imbalance between secretion (too little) and
absorption (relatively too much). The defective salt transport
processes that characterize these mucosal dysfunctions reside in
the epithelial layer of the mucosal surface.
One approach to replenish the protective liquid layer on mucosal
surfaces is to "re-balance" the system by blocking Na.sup.+ channel
and liquid absorption. The epithelial protein that mediates the
rate-limiting step of Na.sup.+ and liquid absorption is the
epithelial Na.sup.+ channel (ENaC). ENaC is positioned on the
apical surface of the epithelium, i.e. the mucosal
surface-environmental interface. Therefore, to inhibit ENaC
mediated Na.sup.+ and liquid absorption, an ENaC blocker of the
amiloride class (which blocks from the extracellular domain of
ENaC) must be delivered to the mucosal surface and, importantly, be
maintained at this site, to achieve therapeutic utility. The
present invention describes diseases characterized by too little
liquid on mucosal surfaces and "topical" sodium channel blockers
designed to exhibit the increased potency, reduced mucosal
absorption, and slow dissociation ("unbinding" or detachment) from
ENaC required for therapy of these diseases.
Chronic obstructive pulmonary diseases are characterized by
dehydration of airway surfaces and the retention of mucous
secretions in the lungs. Examples of such diseases include cystic
fibrosis, chronic bronchitis, and primary or secondary ciliary
dyskinesia. Such diseases affect approximately 15 million patients
in the United States, and are the sixth leading cause of death.
Other airway or pulmonary diseases characterized by the
accumulation of retained mucous secretions include sinusitis (an
inflammation of the paranasal sinuses associated with upper
respiratory infection) and pneumonia.
Chronic bronchitis (CB), including the most common lethal genetic
form of chronic bronchitis, cystic fibrosis (CF), are diseases that
reflect the body's failure to clear mucus normally from the lungs,
which ultimately produces chronic airways infection. In the normal
lung, the primary defense against chronic intrapulmonary airways
infection (chronic bronchitis) is mediated by the continuous
clearance of mucus from bronchial airway surfaces. This function in
health effectively removes from the lung potentially noxious toxins
and pathogens. Recent data indicate that the initiating problem,
i.e., the "basic defect," in both CB and CF is the failure to clear
mucus from airway surfaces. The failure to clear mucus reflects an
imbalance between the amount of liquid and mucin on airway
surfaces. This "airway surface liquid" (ASL) is primarily composed
of salt and water in proportions similar to plasma (i.e.,
isotonic). Mucin macromolecules organize into a well defined "mucus
layer" which normally traps inhaled bacteria and is transported out
of the lung via the actions of cilia which beat in a watery, low
viscosity solution termed the "periciliary liquid" (PCL). In the
disease state, there is an imbalance in the quantities of mucus as
ASL on airway surfaces. This results in a relative reduction in ASL
which leads to mucus concentration, reduction in the lubricant
activity of the PCL, and a failure to clear mucus via ciliary
activity to the mouth. The reduction in mechanical clearance of
mucus from the lung leads to chronic bacterial colonization of
mucus adherent to airway surfaces. It is the chronic retention of
bacteria, the failure of local antimicrobial substances to kill
mucus-entrapped bacteria on a chronic basis, and the consequent
chronic inflammatory responses of the body to this type of surface
infection, that lead to the syndromes of CB and CF.
The current afflicted population in the U.S. is 12,000,000 patients
with the acquired (primarily from cigarette smoke exposure) form of
chronic bronchitis and approximately 30,000 patients with the
genetic form, cystic fibrosis. Approximately equal numbers of both
populations are present in Europe. In Asia, there is little CF but
the incidence of CB is high and, like the rest of the world, is
increasing.
There is currently a large, unmet medical need for products that
specifically treat CB and CF at the level of the basic defect that
cause these diseases. The current therapies for chronic bronchitis
and cystic fibrosis focus on treating the symptoms and/or the late
effects of these diseases. Thus, for chronic bronchitis,
.beta.-agonists, inhaled steroids, anti-cholinergic agents, and
oral theophyllines and phosphodiesterase inhibitors are all in
development. However, none of these drugs treat effectively the
fundamental problem of the failure to clear mucus from the lung.
Similarly, in cystic fibrosis, the same spectrum of pharmacologic
agents is used. These strategies have been complemented by more
recent strategies designed to clear the CF lung of the DNA
("Pulmozyme"; Genentech) that has been deposited in the lung by
neutrophils that have futilely attempted to kill the bacteria that
grow in adherent mucus masses and through the use of inhaled
antibiotics ("TOBI") designed to augment the lungs' own killing
mechanisms to rid the adherent mucus plaques of bacteria. A general
principle of the body is that if the initiating lesion is not
treated, in this case mucus retention/obstruction, bacterial
infections became chronic and increasingly refractory to
antimicrobial therapy. Thus, a major unmet therapeutic need for
both CB and CF lung diseases is an effective means of re-hydrating
airway mucus (i.e., restoring/expanding the volume of the ASL) and
promoting its clearance, with bacteria, from the lung.
R. C. Boucher, in U.S. Pat. No. 6,264,975, describes the use of
pyrazinoylguanidine sodium channel blockers for hydrating mucosal
surfaces. These compounds, typified by the well-known diuretics
amiloride, benzamil, and phenamil, are effective. However, these
compounds suffer from the significant disadvantage that they are
(1) relatively impotent, which is important because the mass of
drug that can be inhaled by the lung is limited; (2) rapidly
absorbed, which limits the half-life of the drug on the mucosal
surface; and (3) are freely dissociable from ENaC. The sum of these
disadvantages embodied in these well known diuretics produces
compounds with insufficient potency and/or effective half-life on
mucosal surfaces to have therapeutic benefit for hydrating mucosal
surfaces.
R. C. Boucher, in U.S. Pat. No. 6,926,911, suggests the use of the
relatively impotent sodium channel blockers such as amiloride, with
osmolytes for the treatment of airway diseases. This combination
gives no practical advantage over either treatment alone and is
clinically not useful, see Donaldson et al, N Eng J. Med., 2006;
353:241-250. Amiloride was found to block the water permeability of
airways and negate the potential benefit of concurrent use of
hypertonic saline and amiloride.
U.S. Pat. No. 5,817,028 to Anderson describes a method for the
provocation of air passage narrowing (for evaluating susceptibility
to asthma) and/or the induction of sputum in subjects via the
inhalation of mannitol. It is suggested that the same technique can
be used to induce sputum and promote mucociliary clearance.
Substances suggested include sodium chloride, potassium chloride,
mannitol and dextrose.
Clearly, what is needed are drugs that are more effective at
restoring the clearance of mucus from the lungs of patients with
CB/CF. The value of these new therapies will be reflected in
improvements in the quality and duration of life for both the CF
and the CB populations.
Other mucosal surfaces in and on the body exhibit subtle
differences in the normal physiology of the protective surface
liquids on their surfaces but the pathophysiology of disease
reflects a common theme, i.e., too little protective surface
liquid. For example, in xerostomia (dry mouth) the oral cavity is
depleted of liquid due to a failure of the parotid sublingual and
submandibular glands to secrete liquid despite continued Na.sup.+
(ENaC) transport mediated liquid absorption from the oral cavity.
Similarly, keratoconjunctivitis sira (dry eye) is caused by failure
of lacrimal glands to secrete liquid in the face of continued
Na.sup.+ dependent liquid absorption on conjunctional surfaces. In
rhinosinusitis, there is an imbalance, as in CB, between mucin
secretion and relative ASL depletion. Finally, in the
gastrointestinal tract, failure to secrete Cl-- (and liquid) in the
proximal small intestine, combined with increased Na.sup.+ (and
liquid) absorption in the terminal ileum leads to the distal
intestinal obstruction syndrome (DIOS). In older patients excessive
Na.sup.+ (and volume) absorption in the descending colon produces
constipation and diverticulitis.
Fifty million Americans and hundreds of millions of others around
the world suffer from high blood pressure and the subsequent
sequale leading to congestive heart failure and increasing
mortality. It is the Western World's leading killer and there is a
need there for new medicines to treat these diseases. Thus, in
addition, some of the novel sodium channel blockers of this
invention can be designed to target the kidney and as such they may
be used as diuretics for the treatment of hypertension, congestive
heart failure (CHF) and other cardiovascular diseases. These new
agents may be used alone or in combination with beta-blockers, ACE
inhibitors, HMGCoA reductase inhibitors, calcium channel blockers
and other cardiovascular agents.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide compounds that
are more potent and/or absorbed less rapidly from mucosal surfaces,
and/or are less reversible as compared to known compounds.
It is another aspect of the present invention to provide compounds
that are more potent and/or absorbed less rapidly and/or exhibit
less reversibility, as compared to compounds such as amilorde,
benzamil, and phenamil. Therefore, the compounds will give a
prolonged pharmacodynamic half-life on mucosal surfaces as compared
to known compounds.
It is another object of the present invention to provide compounds
which are (1) absorbed less rapidly from mucosal surfaces,
especially airway surfaces, as compared to known compounds and; (2)
when absorbed from musosal surfaces after administration to the
mucosal surfaces, are converted in vivo into metabolic derivitives
thereof which have reduced efficacy in blocking sodium channels as
compared to the administered parent compound. It is another object
of the present invention to provide compounds that are more potent
and/or absorbed less rapidly and/or exhibit less reversibility, as
compared to compounds such as amiloride, benzamil, and phenamil.
Therefore, such compounds will give a prolonged pharmacodynamic
half-life on mucosal surfaces as compared to previous
compounds.
It is another object of the present invention to provide compounds
that target the kidney for use in the treatment of cardiovascular
disease.
It is another object of the present invention to provide methods of
treatment that take advantage of the pharmacological properties of
the compounds described above.
In particular, it is an object of the present invention to provide
methods of treatment which rely on rehydration of mucosal
surfaces.
In particular, it is an object of the present invention to provide
methods of treating cardiovascular disease.
The objects of the present invention may be accomplished with a
class of pyrazinoylguanidine represented by a compound of formula
(I)
##STR00002## and includes racemates, enantiomers, diastereomers,
tautomers, polymorphs, pseudopolymorphs and pharmaceutically
acceptable salts, thereof, wherein:
X is hydrogen, halogen, trifluoromethyl, lower alkyl, unsubstituted
or substituted phenyl, lower alkyl-thio, phenyl-lower alkyl-thio,
lower alkyl-sulfonyl, or phenyl-lower alkyl-sulfonyl;
Y is hydrogen, hydroxyl, mercapto, lower alkoxy, lower alkyl-thio,
halogen, lower alkyl, unsubstituted or substituted mononuclear
aryl, or --N(R.sup.2).sub.2;
R.sup.1 is hydrogen or lower alkyl;
each R.sup.2 is, independently, --R.sup.7,
--(CH.sub.2).sub.m--OR.sup.8, --(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7, or
##STR00003##
R.sup.3 and R.sup.4 are each, independently, hydrogen, lower alkyl,
hydroxyl-lower alkyl, phenyl, (phenyl)-lower alkyl,
(halophenyl)-lower alkyl, ((lower-alkyl)phenyl)-lower-alkyl,
((lower-alkoxy)phenyl))-lower alkyl, (naphthyl)-lower alkyl, or
(pyridyl)-lower alkyl, or a group represented by formula A or
formula B, with the proviso that at least one of R.sup.3 and
R.sup.4 is a group represented by the formula A or formula B;
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1
formula A:
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2
formula B:
A.sup.1 is a C.sub.7-C.sub.15-membered aromatic carbocycle
substituted with at least one R.sup.5 and the remaining
substituents are R.sup.6;
A.sup.2 is a seven to fifteen-membered aromatic heterocycle
substituted with at least one R.sup.5 and the remaining
substituents are R.sup.6 wherein said aromatic heterocycle
comprises 1-4 heteroatoms selected from the group consisting of O,
N, and S;
each R.sup.L is, independently, --R.sup.7,
--(CH.sub.2).sub.n--OR.sup.8, --O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00004##
each o is, independently, an integer from 0 to 10;
each p is, independently, an integer from 0 to 10;
with the proviso that the sum of o and p in each contiguous chain
is from 1 to 10;
each x is, independently, O, NR.sup.10, C(.dbd.O), CHOH,
C(.dbd.N--R.sup.10), CHNR.sup.7R.sup.10, or a single bond;
each R.sup.5 is, independently, OH, --(CH.sub.2).sub.m--OR.sup.8,
--O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00005## --(CH.sub.2).sub.n--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CO.sub-
.2R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CO.-
sub.2R.sup.13, --(CH.sub.2).sub.n--(CHOR.sup.8).sub.m,
--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m(Z).sub.g--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.m--(Z).sub.g--CO.sub.2R.sup-
.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CO.s-
ub.2R.sup.13,
--(CH.sub.2).sub.n--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--CO--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13-
,
-Het-(CH.sub.2).sub.n--(Z).sub.g--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.su-
p.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C-
ONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m--(CHOR.sup.8).sub.n--C-
ONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONH--C(.dbd.NR.sup.13)--NR.sup.1-
3R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONH--C(.dbd.NR.sup.-
13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.dbd.NR.sup.13-
)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.dbd.NR.sup-
.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONH--C(.dbd.NR.sup.13)--N-
R.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONH--C(.dbd.NR.sup.13)-
--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.db-
d.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.-
dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--CONR.sup.7--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7--CONR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7--CONR.sup.13R.sup.13-
,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7--CONR.sup.-
13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CNR.sup.7--CONR.sup-
.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7--CONR.sup.13R.s-
up.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7--CONR.-
sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.n--(Z).sub.g--CONR.sup.7--C-
ONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7-
--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7SO.sub.2NR.sup.13R.sup.-
13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.7SO.sub.2NR.sup.13-
R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7SO.-
sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7SO.sub.2N-
R.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7SO.sub.2NR.sup.1-
3R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7SO.-
sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7S-
O.sub.2NR.sup.13R.sup.13,
--Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.su-
p.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--SO.sub-
.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--SO.-
sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2NR.sup.13R.sup-
.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2NR.sup.-
13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mSO.sub.2NR.sup.13R.sup.13,
--Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mSO.sub.2NR.sup.13R.sup-
.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.-
2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2-
NR.sup.13R.sup.13, --(CH.sub.2).sub.n--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13R.sup.13-
,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13-
R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--
-CONR.sup.13R.sup.13, --(CH.sub.2).sub.n--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7COR.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7COR.sup.1-
3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup-
.7COR.sup.13, --(CH.sub.2).sub.n--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--,
--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7CO.sub.2R.su-
p.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7CO.s-
ub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7CO.sub.2R.sup.13-
,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7CO.sub.2R.su-
p.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.s-
up.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup-
.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--NH---
C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--NH--
-C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R-
.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--NH--C(.dbd.NR.sup.13)--
-NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--
-NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.1-
3)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.-
NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(NR-
.sup.13)--NR.sup.13R.sup.13,
CH.sub.2).sub.n--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--C(.dbd.NH)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--C(.dbd.NH)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--(Z).sub.g--C(.dbd.NH)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C(.dbd-
.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C(.d-
bd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--C(.dbd.NR.sup.13)--NR.sup.13R.sup-
.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--C(.dbd.NR.sup.13)--NR.sup.-
13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
--Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup.13)--
-NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--C(.dbd.NHC(.dbd.NR.sup.1-
3)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup-
.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.N-
R.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
--(CH.sub.2).sub.nNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--(CH.sub.2).sub.n--N.sup..sym.--(R.sup.11).sub.3,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
--(CH.sub.2).sub.n--(C.dbd.O)NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--(C.dbd.O)NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--(CHOR.sup.8).sub.mCH.sub.2NR.sup.10--(Z).sub.g--R.-
sup.10,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.10--(Z).sub-
.g--R.sup.10,
--(CH.sub.2).sub.nNR.sup.10--O(CH.sub.2).sub.m(CHOR.sup.8).sub.nCH.sub.2N-
R.sup.10--(Z).sub.g--R.sup.10,
--O(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m--(CHOR.sup.8).sub.nCH.su-
b.2NR.sup.10--(Z).sub.g--R.sup.10,
-(Het)-(CH.sub.2).sub.m--OR.sup.8,
-(Het)-(CH.sub.2).sub.m--NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
-(Het)-(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.-
n--CH.sub.2OR.sup.8, -(Het)-(CH.sub.2).sub.m--CO.sub.2R.sup.7,
-(Het)-(CH.sub.2).sub.m--NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.n--NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(Z).sub.gR.sup.12,
-(Het)-(CH.sub.2).sub.mNR.sup.11R.sup.11,
-(Het)-(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
-(Het)-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(C.dbd.O)NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(CHOR.sup.8).sub.mCH.sub.2NR.sup.10--(Z).sub.g---
R.sup.10,
-(Het)-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m--(CHOR.sup.-
8).sub.nCH.sub.2NR.sup.10--(Z).sub.g--R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, Link-(CH.sub.2).sub.n-CAP,
Link-(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2-CAP,
Link-(CH.sub.2).sub.n--(Z).sub.g-CAP,
Link-(CH.sub.2).sub.n(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--NR.sup.13--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
CAP,
Link-(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.13--(Z).sub-
.g-CAP,
Link-(CH.sub.2).sub.nNR.sup.13--(CH.sub.2).sub.m(CHOR.sup.8).sub.n-
CH.sub.2NR.sup.13--(Z).sub.g-CAP,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-NH--C(.dbd.O)--NH--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m--C(.dbd.O)NR.-
sup.10R.sup.10,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.11R.sup.11,
Link-(CH.sub.2).sub.mC(.dbd.O)NR.sup.12R.sup.12,
Link-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--(Z).sub.g-CAP,
Link-(Z).sub.g--(CH.sub.2).sub.m-Het-(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CR.sup.11R.sup.11-CA-
P, Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n--(Z).sub.g--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n(Z).sub.g--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n--NR.sup.13--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.13--(Z).sub.g---
CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.nNR.sup.13--(CH.sub.2).sub.m(CHOR.sup.8).sub.nCH.sub.-
2NR.sup.13--(Z).sub.g--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP-
, Link NH--C(.dbd.O)--NH--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m--CR.sup.11R.s-
up.11-CAP,
Link-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--(Z).sub.g---
CR.sup.11R.sup.11-CAP, or
Link-(Z).sub.g--(CH.sub.2).sub.m-Het-(CH.sub.2).sub.m--CR.sup.11R.sup.11--
CAP;
each R.sup.6 is, independently, R.sup.5, --R.sup.7, --OR.sup.11,
--N(R.sup.7).sub.2, --(CH.sub.2).sub.m--OR.sup.8,
--O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00006##
wherein when two R.sup.6 are --OR.sup.11 and are located adjacent
to each other on the aromatic carbocycle or aromatic heterocycle,
the two OR.sup.11 may form a methylenedioxy group;
each R.sup.7 is, independently, hydrogen, lower alkyl, phenyl,
substituted phenyl or
--CH.sub.2(CHOR.sup.8).sub.m--CH.sub.2OR.sup.8;
each R.sup.8 is, independently, hydrogen, lower alkyl,
--C(.dbd.O)--R.sup.11, glucuronide, 2-tetrahydropyranyl, or
##STR00007##
each R.sup.9 is, independently, --CO.sub.2R.sup.7,
--CON(R.sup.7).sub.2, --SO.sub.2CH.sub.3, --C(.dbd.O)R.sup.7,
--CO.sub.2R.sup.13, --CON(R.sup.13).sub.2,
--SO.sub.2CH.sub.2R.sup.13, or --C(.dbd.O)R.sup.13;
each R.sup.10 is, independently, --H, --SO.sub.2CH.sub.3,
--CO.sub.2R.sup.7, --C(.dbd.O)NR.sup.7R.sup.9, --C(.dbd.O)R.sup.7,
or --CH.sub.2--(CHOH).sub.n--CH.sub.2OH;
each Z is, independently, --(CHOH)--, --C(.dbd.O)--,
--(CHNR.sup.7R.sup.10)--, --(C.dbd.NR.sup.10)--, --NR.sup.10--,
--(CH.sub.2).sub.n--, --(CHNR.sup.13R.sup.13)--,
--(C.dbd.NR.sup.13)--, or --NR.sup.13--;
each R.sup.11 is, independently, hydrogen, lower alkyl, phenyl
lower alkyl or substituted phenyl lower alkyl;
each R.sup.12 is, independently, --SO.sub.2CH.sub.3,
--CO.sub.2R.sup.7, --C(.dbd.O)NR.sup.7R.sup.9, --C(.dbd.O)R.sup.7,
--CH.sub.2(CHOH).sub.n--CH.sub.2OH, --CO.sub.2R.sup.13,
--C(.dbd.O)NR.sup.13R.sup.13, or --C(.dbd.O)R.sup.13;
each R.sup.13 is, independently, R.sup.7, R.sup.10,
--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.m--NR.sup.7R.sup.7,
--(CH.sub.2).sub.m--NR.sup.11R.sup.11R.sup.11).sup.+,
--(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--(CH.sub.2).sub.mNR.sup.11R.sup.11-
,
--(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--(CH.sub.2).sub.mNR.sup.7R.sup.10-
, --(CH.sub.2).sub.m--NR.sup.10R.sup.10,
--(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--(CH.sub.2).sub.m--(NR.sup.11R.sup-
.11R.sup.11).sup.+,
--(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--(CH.sub.2).sub.mNR.sup.7R.sup.7,
##STR00008##
with the proviso that in the moiety --NR.sup.13R.sup.13, the two
R.sup.13 along with the nitrogen to which they are attached may,
optionally, form a ring selected from:
##STR00009##
each V is, independently, --(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.m--, --NR.sup.7R.sup.7,
--(CH.sub.2).sub.m--(NR.sup.11R.sup.11R.sup.11).sup.+,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(CH.sub.2).sub.mNR.sup.7R.sup.10,
--(CH.sub.2).sub.n--NR.sup.10R.sup.10--(CH.sub.2).sub.n--(CHOR.sup.8).sub-
.m--(CH.sub.2).sub.mNR.sup.7R.sup.7,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(CH.sub.2).sub.m--(NR.sup.11R.sup-
.11R.sup.11).sup.+ with the proviso that when V is attached
directly to a nitrogen atom, then V can also be, independently,
R.sup.7, R.sup.10, or (R.sup.11).sub.2;
each R.sup.14 is, independently, H, R.sup.12,
--(CH.sub.2).sub.n--SO.sub.2CH.sub.3,
--(CH.sub.2).sub.n--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--C(.dbd.O)R.sup.13,
--(CH.sub.2).sub.n--(CHOH).sub.n--CH.sub.2OH,
--NH--(CH.sub.2).sub.n--SO.sub.2CH.sub.3,
NH--(CH.sub.2).sub.n--C(.dbd.O)R.sup.11,
NH--C(.dbd.O)--NH--C(.dbd.O)R.sup.11, --C(.dbd.O)NR.sup.13R.sup.13,
--OR.sup.11, --NH--(CH.sub.2).sub.n--R.sup.10, --Br, --Cl, --F,
--I, SO.sub.2NHR.sup.11, --NHR.sup.13,
--NH--C(.dbd.O)--NR.sup.13R.sup.13, --(CH.sub.2).sub.n--NHR.sup.13,
or --NH--(CH.sub.2).sub.n--C(.dbd.O)--R.sup.13;
each g is, independently, an integer from 1 to 6; each m is,
independently, an integer from 1 to 7;
each n is, independently, an integer from 0 to 7;
each -Het- is, independently, --N(R.sup.7)--, --N(R.sup.10)--,
--S--, --SO--, --SO.sub.2--; --O--, --SO.sub.2NH--, --NHSO.sub.2--,
--NR.sup.7CO--, --CONR.sup.7--, --N(R.sup.13)--,
--SO.sub.2NR.sup.13--, --NR.sup.13CO--, or --CONR.sup.13--;
each Link is, independently, --O--, --(CH.sub.2).sub.n--,
--O(CH.sub.2).sub.m--, --NR.sup.13--C(.dbd.O)--NR.sup.13--,
--NR.sup.13--C(.dbd.O)--(CH.sub.2).sub.m--,
--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m.sup.-,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.n--, --S--, --SO--,
--SO.sub.2--, --SO.sub.2NR.sup.7--, --SO.sub.2NR.sup.10--, or
-Het-;
each CAP is, independently, thiazolidinedione, oxazolidinedione,
-heteroaryl-C(.dbd.O)N R.sup.13R.sup.13, heteroaryl-W, --CN,
--O--C(.dbd.S)NR.sup.13R.sup.13, --(Z).sub.gR.sup.13,
--CR.sup.10((Z).sub.gR.sup.13)((Z).sub.gR.sup.13), --C(.dbd.O)OAr,
--C(.dbd.O)NR.sup.13Ar, imidazoline, tetrazole, tetrazole amide,
--SO.sub.2NHR.sup.13,
--SO.sub.2NH--C(R.sup.13R.sup.13)--(Z).sub.g--R.sup.13, a cyclic
sugar or oligosaccharide, a cyclic amino sugar, oligosaccharide,
--CR.sup.10(--(CH.sub.2).sub.m--R.sup.9)(--(CH.sub.2).sub.m--R.sup.9),
--N(--(CH.sub.2).sub.m--R.sup.9)(--(CH.sub.2).sub.m--R.sup.9),
--NR.sup.13(--(CH.sub.2).sub.m--CO.sub.2R.sup.13),
##STR00010##
each Ar is, independently, phenyl, substituted phenyl, wherein the
substituents of the substituted phenyl are 1-3 substituents
independently selected from the group consisting of OH, OCH.sub.3,
NR.sup.13R.sup.13, Cl, F, and CH.sub.3, or heteroaryl; and
each W is, independently, thiazolidinedione, oxazolidinedione,
heteroaryl-C(.dbd.O)N R.sup.13R.sup.13, --CN,
--O--C(.dbd.S)NR.sup.13R.sup.13, --(Z).sub.gR.sup.13,
--CR.sup.10((Z).sub.gR.sup.13)((Z).sub.gR.sup.13), --C(.dbd.O)OAr,
--C(.dbd.O)N R.sup.13Ar, imidazoline, tetrazole, tetrazole amide,
--SO.sub.2NHR.sup.13,
--SO.sub.2NH--C(R.sup.13R.sup.13)--(Z).sub.g--R.sup.13, a cyclic
sugar or oligosaccharide, a cyclic amino sugar,
oligosaccharide,
##STR00011##
with the proviso that when any --CHOR.sup.8-- or --CH.sub.2OR.sup.8
groups are located 1,2- or 1,3- with respect to each other, the
R.sup.8 groups may, optionally, be taken together to form a cyclic
mono- or di-substituted 1,3-dioxane or 1,3-dioxolane.
The present also provides pharmaceutical compositions which
comprise a compound described herein.
The present invention also provides a method of promoting hydration
of mucosal surfaces, comprising:
administering an effective amount of a compound described herein to
a mucosal surface of a subject.
The present invention also provides a method of restoring mucosal
defense, comprising:
topically administering an effective amount of compound described
herein to a mucosal surface of a subject in need thereof.
The present invention also provides a method of blocking ENaC,
comprising:
contacting sodium channels with an effective amount of a compound
represented by described herein.
The present invention also provides a method of promoting mucus
clearance in mucosal surfaces, comprising:
administering an effective amount of a compound represented
described herein to a mucosal surface of a subject.
The present invention also provides a method of treating chronic
bronchitis, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating cystic
fibrosis, comprising:
administering an effective amount of compound described herein to a
subject in need thereof.
The present invention also provides a method of treating
rhinosinusitis, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating nasal
dehydration, comprising:
administering an effective amount of a compound described herein to
the nasal passages of a subject in need thereof.
In a specific embodiment, the nasal dehydration is brought on by
administering dry oxygen to the subject.
The present invention also provides a method of treating sinusitis,
comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating pneumonia,
comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating
ventilator-induced pneumonia, comprising:
administering an effective compound described herein to a subject
by means of a ventilator.
The present invention also provides a method of treating asthma,
comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating primary
ciliary dyskinesia, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating otitis
media, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of inducing sputum for
diagnostic purposes, comprising:
administering an effective amount of compound described herein to a
subject in need thereof.
The present invention also provides a method of treating chronic
obstructive pulmonary disease, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating emphysema,
comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating dry eye,
comprising:
administering an effective amount of a compound described herein to
the eye of the subject in need thereof.
The present invention also provides a method of promoting ocular
hydration, comprising:
administering an effective amount of a compound described herein to
the eye of the subject.
The present invention also provides a method of promoting corneal
hydration, comprising:
administering an effective amount of a compound described herein to
the eye of the subject.
The present invention also provides a method of treating Sjogren's
disease, comprising:
administering an effective amount of compound described herein to a
subject in need thereof.
The present invention also provides a method of treating vaginal
dryness, comprising:
administering an effective amount of a compound described herein to
the vaginal tract of a subject in need thereof.
The present invention also provides a method of treating dry skin,
comprising:
administering an effective amount of a compound described herein to
the skin of a subject in need thereof.
The present invention also provides a method of treating dry mouth
(xerostomia), comprising:
administering an effective amount of compound described herein to
the mouth of the subject in need thereof.
The present invention also provides a method of treating distal
intestinal obstruction syndrome, comprising:
administering an effective amount of compound described herein to a
subject in need thereof.
The present invention also provides a method of treating
esophagitis, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating
bronchiectasis, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating
constipation, comprising:
administering an effective amount of a compound described herein to
a subject in need thereof. In one embodiment of this method, the
compound is administered either orally or via a suppository or
enema.
The present invention also provides a method of treating chronic
diverticulitis comprising:
administering an effective amount of a compound described herein to
a subject in need thereof.
The present invention also provides a method of treating
hypertension, comprising administering the compound described
herein to a subject in need thereof.
The present invention also provides a method of reducing blood
pressure, comprising administering the compound described herein to
a subject in need thereof.
The present invention also provides a method of treating edema,
comprising administering the compound described herein to a subject
in need thereof.
The present invention also provides a method of promoting diuresis,
comprising administering the compound described herein to a subject
in need thereof.
The present invention also provides a method of promoting
natriuresis, comprising administering the compound described herein
to a subject in need thereof.
The present invention also provides a method of promoting
saluresis, comprising administering the compound described herein
to a subject in need thereof.
It is an object of the present invention to provide treatments
comprising the use of osmolytes together with sodium channel
blockers of formula (I) that are more potent, more specific, and/or
absorbed less rapidly from mucosal surfaces, and/or are less
reversible as compared to compounds such as amiloride, benzamil,
and phenamil.
It is another aspect of the present invention to provide treatments
using sodium channel blockers of formula (I) that are more potent
and/or absorbed less rapidly and/or exhibit less reversibility, as
compared to compounds such as amiloride, benzamil, and phenamil
when administered with an osmotic enhancer. Therefore, such sodium
channel blockers when used in conjunction with osmolytes will give
a prolonged pharmacodynamic half-life on mucosal surfaces as
compared to either compound used alone.
It is another object of the present invention to provide treatments
using sodium channel blockers of formula (I) and osmolytes together
which are absorbed less rapidly from mucosal surfaces, especially
airway surfaces, as compared to compounds such as amiloride,
benzamil, and phenamil.
It is another object of the invention to provide compositions which
contain sodium channel blockers of formula (I) and osmolytes.
The objects of the invention may be accomplished with a method of
treating a disease ameliorated by increased mucociliary clearance
and mucosal hydration comprising administering an effective amount
of a compound of formula (I) as defined herein and an osmolyte to a
subject to a subject in need of increased mucociliary clearance and
mucosal hydration.
The objects of the invention may also be accomplished with a method
of inducing sputum for diagnostic purposes, comprising
administering an effective amount of a compound of formula (I) as
defined herein and an osmolyte to a subject in need thereof.
The objects of the invention may also be accomplished with a method
of treating anthrax, comprising administering an effective amount
of a compound of formula (I) as defined herein and an osmolyte to a
subject in need thereof.
The objects of the invention may also be accomplished with a method
of prophylactic, post-exposure prophylactic, preventive or
therapeutic treatment against diseases or conditions caused by
pathogens, particularly pathogens which may be used in
bioterrorism, comprising administering an effective amount of a
compound of formula (I) to a subject in need thereof.
The objects of the invention may also be accomplished with a
composition, comprising a compound of formula (I) as defined herein
and an osmotically active compound.
DETAILED DESCRIPTION OF THE INVENTION
The present invention is based on the discovery that the compounds
of formula (I) are more potent and/or, absorbed less rapidly from
mucosal surfaces, especially airway surfaces, and/or less
reversible from interactions with ENaC as compared to compounds
such as amiloride, benzamil, and phenamil. Therefore, the compounds
of formula (I) have a longer half-life on mucosal surfaces as
compared to these compounds.
The present invention is also based on the discovery that certain
compounds embraced by formula (I) are converted in vivo into
metabolic derivatives thereof that have reduced efficacy in
blocking sodium channels as compared to the parent administered
compound, after they are absorbed from mucosal surfaces after
administration. This important property means that the compounds
will have a lower tendency to cause undesired side-effects by
blocking sodium channels located at untargeted locations in the
body of the recipient, e.g., in the kidneys.
The present invention is also based on the discovery that certain
compounds embraced by formula (1) target the kidney and thus may be
used as cardiovascular agents.
In the compounds represented by formula (I), X may be hydrogen,
halogen, trifluoromethyl, lower alkyl, lower cycloalkyl,
unsubstituted or substituted phenyl, lower alkyl-thio, phenyl-lower
alkyl-thio, lower alkyl-sulfonyl, or phenyl-lower alkyl-sulfonyl.
Halogen is preferred.
Examples of halogen include fluorine, chlorine, bromine, and
iodine. Chlorine and bromine are the preferred halogens. Chlorine
is particularly preferred. This description is applicable to the
term "halogen" as used throughout the present disclosure.
As used herein, the term "lower alkyl" means an alkyl group having
less than 8 carbon atoms. This range includes all specific values
of carbon atoms and subranges there between, such as 1, 2, 3, 4, 5,
6, and 7 carbon atoms. The term "alkyl" embraces all types of such
groups, e.g., linear, branched, and cyclic alkyl groups. This
description is applicable to the term "lower alkyl" as used
throughout the present disclosure. Examples of suitable lower alkyl
groups include methyl, ethyl, propyl, cyclopropyl, butyl, isobutyl,
etc.
Substituents for the phenyl group include halogens. Particularly
preferred halogen substituents are chlorine and bromine.
Y may be hydrogen, hydroxyl, mercapto, lower alkoxy, lower
alkyl-thio, halogen, lower alkyl, lower cycloalkyl, mononuclear
aryl, or --N(R.sup.2).sub.2. The alkyl moiety of the lower alkoxy
groups is the same as described above. Examples of mononuclear aryl
include phenyl groups. The phenyl group may be unsubstituted or
substituted as described above. The preferred identity of Y is
--N(R.sup.2).sub.2. Particularly preferred are such compounds where
each R.sup.2 is hydrogen.
R.sup.1 may be hydrogen or lower alkyl. Hydrogen is preferred for
R.sup.1.
Each R.sup.2 may be, independently, --R.sup.7,
--(CH.sub.2).sub.m--OR.sup.8, --(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7, or
##STR00012##
Hydrogen and lower alkyl, particularly C.sub.1-C.sub.3 alkyl, are
preferred for R.sup.2. Hydrogen is particularly preferred.
R.sup.3 and R.sup.4 may be, independently, hydrogen, lower alkyl,
hydroxyl-lower alkyl, phenyl, (phenyl)-lower alkyl,
(halophenyl)-lower alkyl, ((lower-alkyl)phenyl)-lower-alkyl),
(lower-alkoxyphenyl)-lower alkyl, (naphthyl)-lower alkyl,
(pyridyl)-lower alkyl or a group represented by
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1 or
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2,
provided that at least one of R.sup.3 and R.sup.4 is a group
represented by
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1 or
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2.
Preferred compounds are those where one of R.sup.3 and R.sup.4 is
hydrogen and the other is represented by
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1 or
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2. In a
particularly preferred aspect one of R.sup.3 and R.sup.4 is
hydrogen and the other of R.sup.3 or R.sup.4 is represented by
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1. In
another particularly preferred aspect one of R.sup.3 and R.sup.4 is
hydrogen and the other of R.sup.3 or R.sup.4 is represented by
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2.
A moiety --(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.p--
defines an alkylene group bonded to the group A.sup.1 or A.sup.2.
The variables o and p may each, independently, be an integer from 0
to 10, subject to the proviso that the sum of o and p in the chain
is from 1 to 10. Thus, o and p may each be 0, 1, 2, 3, 4, 5, 6, 7,
8, 9, or 10. Preferably, the sum of o and p is from 2 to 6. In a
particularly preferred embodiment, the sum of o and p is 4.
The linking group in the alkylene chain, x, may be, independently,
O, NR.sup.10, C(.dbd.O), CHOH, C(.dbd.N--R.sup.10),
CHNR.sup.7R.sup.10, or a single bond;
Therefore, when x is a single bond, the alkylene chain bonded to
the ring is represented by the formula
--(C(R.sup.L).sub.2).sub.o+p--, in which the sum o+p is from 1 to
10.
Each R.sup.L may be, independently, --R.sup.7,
--(CH.sub.2).sub.n--OR.sup.8, --O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00013##
The term --O-glucuronide, unless otherwise specified, means a group
represented by
##STR00014## wherein the O means the glycosidic linkage can be
above or below the plane of the ring.
The term --O-glucose, unless otherwise specified, means a group
represented by
##STR00015## wherein the O means the glycosidic linkage can be
above or below the plane of the ring.
The preferred R.sup.L groups include --H, --OH, --N(R.sup.7).sub.2,
especially where each R.sup.7 is hydrogen.
In the alkylene chain in
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.1 or
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.pA.sup.2, it is
preferred that when one R.sup.L group bonded to a carbon atoms is
other than hydrogen, then the other R.sup.L bonded to that carbon
atom is hydrogen, i.e., the formula --CHR.sup.L--. It is also
preferred that at most two R.sup.L groups in an alkylene chain are
other than hydrogen, wherein the other R.sup.L groups in the chain
are hydrogens. Even more preferably, only one R.sup.L group in an
alkylene chain is other than hydrogen, wherein the other R.sup.L
groups in the chain are hydrogens. In these embodiments, it is
preferable that x is a single bond.
In another particular embodiment of the invention, all of the
R.sup.L groups in the alkylene chain are hydrogen. In these
embodiments, the alkylene chain is represented by the formula
--(CH.sub.2).sub.o-x-(CH.sub.2).sub.p--.
A.sup.1 is a C.sub.7-C.sub.15-membered aromatic carbocycle
substituted with at least one R.sup.5 and the remaining
substituents are R.sup.6. The term aromatic is well known term of
chemical art and designates conjugated systems of 4n'+2 electrons
that are within a ring system, that is with 6, 10, 14, etc.
.pi.-electrons wherein, according to the rule of Huckel, n' is 1,
2, 3, etc. The 4n'+2 electrons may be in any size ring including
those with partial saturation so long as the electrons are
conjugated. For instance, but not by way of limitation,
5H-cyclohepta-1,3,5-triene, benzene, naphthalene,
1,2,3,4-tetrahydronaphthalene etc. would all be considered
aromatic.
The C.sub.7-C.sub.15 aromatic carbocycle may be monocyclic,
bicyclic, or tricyclic and may include partially saturated rings.
Non-limiting examples of these aromatic carbocycles comprise
5H-cyclohepta-1,3,5-triene, naphthalene, phenanthrene, azulene,
anthracene. 1,2,3,4-tetrahydronapthalene, 1,2-dihydronapthalene,
indene, 5H-dibenzo[a,d]cycloheptene, etc.
The C.sub.7-C.sub.15 aromatic carbocycle may be attached to the
--(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.p-- moiety
through any ring carbon atom as appropriate, unless otherwise
specified. Therefore, when partially saturated bicyclic aromatic is
1,2-dihydronapthalene, it may be 1,2-dihydronapthalen-1-yl,
1,2-dihydronapthalen-3-yl, 1,2-dihydronapthalen-5-yl, etc. In a
preferred embodiment A.sup.1 is indenyl, napthalenyl,
1,2-dihydronapthalenyl, 1,2,3,4-tetrahydronapthalenyl, anthracenyl,
fluorenyl, phenanthrenyl, azulenyl, cyclohepta-1,3,5-trienyl or
5H-dibenzo[a,d]cycloheptenyl. In another preferred embodiment,
A.sup.1 is napthalen-1-yl. In another preferred embodiment, A.sup.1
is napthalen-2-yl.
In another preferred embodiment, A.sup.1 is
##STR00016## wherein each Q is, independently, C--H, C--R.sup.5, or
C--R.sup.6, with the proviso that at least one Q is C--R.sup.5.
Therefore, Q may be 1, 2, 3, 4, 5, or 6 C--H. Therefore, Q may be
1, 2, 3, 4, 5, or 6 C--R.sup.6. In a particularly preferred
embodiment, each R.sup.6 is H.
In another preferred embodiment, A.sup.1 is
##STR00017## wherein each Q is, independently, C--H, C--R.sup.5,
C--R.sup.6, with the proviso that at least one Q is C--R.sup.5.
Therefore, Q may be 1, 2, 3, 4, 5, or 6 C--H. Therefore, Q may be
1, 2, 3, 4, 5, or 6 C--R.sup.6. In a particularly preferred
embodiment, each R.sup.6 is H.
In a particularly preferred embodiment, A.sup.1 is
##STR00018##
In another particularly preferred embodiment, A.sup.1 is
##STR00019##
A.sup.2 is a seven to fifteen-membered aromatic heterocycle
substituted with at least one R.sup.5 and the remaining
substituents are R.sup.6 wherein the aromatic heterocycle comprises
1-4 heteroatoms selected from the group consisting of O, N, and
S.
The seven to fifteen-membered aromatic heterocycle may be
monocyclic, bicyclic, or tricyclic and may include partially
saturated rings. Non limiting examples of these aromatic
heterocycles include 1H-azepine, benzo[b]furan, benzo[b]thiophene,
isobenzofuran, isobenzothiophene, 2,3-dihydrobenzo[b]furan,
benzo[b]thiophene, 2,3-dihydrobenzo[b]thiophene, indolizine,
indole, isoindole benzoxazole, benzimidazole, indazole,
benzisoxazole, benzisothizole, benzopyrazole, benzoxadiazole,
benzothiadiazole, benzotriazole, purine, quinoline,
1,2,3,4-tetrahydroquinoline, 3,4-dihydro-2H-chromene,
3,4-dihydro-2H-thiochromene, isoquinoline, cinnoline, quinolizine,
phthalazine, quinoxaline, quinazoline, naphthiridine, pteridine,
benzopyrane, pyrrolopyridine, pyrrolopyrazine, imidazopyrdine,
pyrrolopyrazine, thienopyrazine, furopyrazine, isothiazolopyrazine,
thiazolopyrazine, isoxazolopyrazine, oxazolopyrazine,
pyrazolopyrazine, imidazopyrazine, pyrrolopyrimidine,
thienopyrimidine, furopyrimidine, isothiazolopyrimidine,
thiazolopyrimidine, isoxazolopyrimidine, oxazolopyrimidine,
pyrazolopyrimidine, imidazopyrimidine, pyrrolopyridazine,
thienopyridazine, furopyridazine, isothiazolopyridazine,
thiazolopyridazine, oxazolopyridazine, thiadiazolopyrazine,
oxadiazolopyrimidine, thiadiazolopyrimidine, oxadiazolopyridazine,
thiazolopyridazine, imidazooxazole, imidazothiazole,
imidazoimidazole, isoxazolotriazine, isothiazolotriazine,
oxazolotriazine, thiazolotriazine, carbazole, acridine, phenazine,
phenothiazine, phenooxazine, and 5H-dibenz[b,f]azepine,
10,11-dihydro-5H-dibenz[b,f]azepine, etc.
The seven to fifteen-membered aromatic heterocycle may be attached
to the --(C(R.sup.L).sub.2).sub.o-x-(C(R.sup.L).sub.2).sub.p--
moiety through any ring carbon atom or ring nitrogen atom so long
as a quanternary nitrogen atom is not formed by the attachment.
Therefore, when partially saturated aromatic heterocycle is
1H-azepine, it may be 1H-azepin-1-yl, 1H-azepin-2-yl,
1H-azepin-3-yl, etc. Preferred aromatic heterocycles are
indolizinyl, indolyl, isoindolyl, indolinyl, benzo[b]furanyl,
2,3-dihydrobenzo[b]furanyl, benzo[b]thiophenyl,
2,3-dihydrobenzo[b]thiophenyl, indazolyl, benzimidazolyl,
benzthiazolyl, purinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl.
3,4-dihydro-2H-chromenyl, 3,4-dihydro-2H-thiochromenyl,
isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl,
quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl,
acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl,
dibenzofuranyl, dibenzothiophenyl, 1H-azepinyl,
5H-dibenz[b,f]azepinyl, are
10,11-dihydro-5H-dibenz[b,f]azepinyl.
In another preferred embodiment, A.sup.2 is
##STR00020## wherein each Q is, independently, C--H, C--R.sup.5,
C--R.sup.6, or a nitrogen atom, with the proviso that at least one
Q is nitrogen and one Q is C--R.sup.5, and at most three Q in a
ring are nitrogen atoms. Therefore, in any one ring, each Q may be
1, 2, or 3 nitrogen atoms. In a preferred embodiment, only one Q in
each ring is nitrogen. In another preferred embodiment, only a
single Q is nitrogen. Optionally, 1, 2, 3, 4, or 5 Q may be
C--R.sup.6. Optionally, Q may be 1, 2, 3, 4, or 5 C--H. In a
particularly preferred embodiment, each R.sup.6 is H.
In another preferred embodiment, A.sup.2 is
##STR00021## wherein each Q is, independently, C--H, C--R.sup.5,
C--R.sup.6, or a nitrogen atom, with the proviso that at least one
Q is nitrogen and one Q is C--R.sup.5, and at most three Q in a
ring are nitrogen atoms. Therefore, in any one ring, each Q may be
1, 2, or 3 nitrogen atoms. In a preferred embodiment, only one Q in
each ring is nitrogen. In another preferred embodiment, only a
single Q is nitrogen. Optionally, Q may be 1, 2, 3, 4, or 5 C--H.
Optionally, 1, 2, 3, 4, or 5 Q may be C--R.sup.6. In a particularly
preferred embodiment, each R.sup.6 is H.
Each R.sup.5 is, independently, OH, --(CH.sub.2).sub.m--OR.sup.8,
--O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00022## --(CH.sub.2).sub.n--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CO.sub-
.2R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CO.-
sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m(Z).sub.g--CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m(Z).sub.g--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.m--(Z).sub.g--CO.sub.2R.sup-
.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CO.s-
ub.2R.sup.13,
--(CH.sub.2).sub.n--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--CO--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13-
,
-Het-(CH.sub.2).sub.n--(Z).sub.g--CONH--C(.dbd.NR.sup.13)--NR.sup.13R.su-
p.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C-
ONH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
H--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONH--C(.dbd.NR.sup.13)--NR.sup.1-
3R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONH--C(.dbd.NR.sup.-
13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.dbd.NR.sup.13-
)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.dbd.NR.sup-
.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONH--C(.dbd.NR.sup.13)--N-
R.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONH--C(.dbd.NR.sup.13)-
--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.db-
d.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONH--C(.-
dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--CONR.sup.7--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7--CONR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7--CONR.sup.13R.sup.13-
,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7--CONR.sup.-
13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CNR.sup.7--CONR.sup-
.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7--CONR.sup.13R.s-
up.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7--CONR.-
sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7--C-
ONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7-
--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7SO.sub.2NR.sup.13R.sup.-
13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.7SO.sub.2NR.sup.13-
R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7SO.-
sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7SO.sub.2N-
R.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7SO.sub.2NR.sup.1-
3R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7SO.-
sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7S-
O.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup-
.7SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--SO.sub-
.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--SO.-
sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--SO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--SO.sub.2NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2NR.sup.13R.sup-
.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2NR.sup.-
13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mSO.sub.2NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mSO.sub.2NR.sup.13R.sup.-
13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2-
NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--SO.sub.2-
NR.sup.13R.sup.13, --(CH.sub.2).sub.n--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13R.sup.13-
,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.13-
R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--
-CONR.sup.13R.sup.13, --(CH.sub.2).sub.n--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7COR.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m, --CONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7COR.sup.1-
3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7COR.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7COR.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup-
.7COR.sup.13, --(CH.sub.2).sub.n--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--CONR.sup.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CONR.s-
up.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--CON-
R.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--CONR.sup.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--CONR.sup.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7CO.sub.2R.su-
p.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup.7CO.s-
ub.2R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7CO.sub.2R.sup.13-
,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mCONR.sup.7CO.sub.2R.su-
p.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.s-
up.7CO.sub.2R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--CONR.sup-
.7CO.sub.2R.sup.13,
--(CH.sub.2).sub.n--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--NH---
C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--NH--
-C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R-
.sup.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--NH--C(.dbd.NR.sup.13)--
-NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--
-NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.1-
3)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.dbd.-
NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--NH--C(.d-
bd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--C(.dbd.NH)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--C(.dbd.NH)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--(Z).sub.g--C(.dbd.NH)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C(.dbd-
.NR.sup.13)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m(CHOR.sup.8).sub.n--C(.d-
bd.NR.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--C(.dbd.NR.sup.13)--NR.sup.13R.sup-
.13,
-Het-(CH.sub.2).sub.m--(CHOR.sup.8).sub.m--C(.dbd.NR.sup.13)--NR.sup.-
13R.sup.13,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--C(.dbd.NHC(.dbd.NR.sup.1-
3)--NR.sup.13R.sup.13,
Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--C(.dbd.NR.sup.13)--NR.-
sup.13R.sup.13,
(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.NR.sup-
.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CHOR.sup.8).sub.m--(Z).sub.g--C(.dbd.N-
R.sup.13)--NR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
--(CH.sub.2).sub.nNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--(CH.sub.2).sub.n--N.sup..sym.--(R.sup.11).sub.3,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
--(CH.sub.2).sub.n--(C.dbd.O)NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--(C.dbd.O)NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.m--(CHOR.sup.8).sub.mCH.sub.2NR.sup.10--(Z).sub.g--R.-
sup.10,
--(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.10--(Z).sub-
.g--R.sup.10,
--(CH.sub.2).sub.nNR.sup.10--O(CH.sub.2).sub.m(CHOR.sup.8).sub.nCH.sub.2N-
R.sup.10--(Z).sub.g--R.sup.10,
--O(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m--(CHOR.sup.8).sub.nCH.su-
b.2NR.sup.10--(Z).sub.g--R.sup.10,
-(Het)-(CH.sub.2).sub.m--OR.sup.8,
-(Het)-(CH.sub.2).sub.m--NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
-(Het)-(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
-(Het)-(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.-
n--CH.sub.2OR.sup.8, -(Het)-(CH.sub.2).sub.m--CO.sub.2R.sup.7,
-(Het)-(CH.sub.2).sub.m--NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.n--NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(Z).sub.gR.sup.12,
-(Het)-(CH.sub.2).sub.mNR.sup.11R.sup.11,
-(Het)-(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
-(Het)-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-(Het)-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(C.dbd.O)NR.sup.12R.sup.12,
-(Het)-(CH.sub.2).sub.m--(CHOR.sup.8).sub.mCH.sub.2NR.sup.10--(Z).sub.g---
R.sup.10,
-(Het)-(CH.sub.2).sub.m--NR.sup.10--(CH.sub.2).sub.m--(CHOR.sup.-
8).sub.nCH.sub.2NR.sup.10--(Z).sub.g--R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.m--CH.sub.2OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, Link-(CH.sub.2).sub.n-CAP,
Link-(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2-CAP,
Link-(CH.sub.2).sub.n--(Z).sub.g-CAP,
Link-(CH.sub.2).sub.n(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--NR.sup.13--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
CAP,
Link-(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.13--(Z).sub-
.g-CAP,
Link-(CH.sub.2).sub.nNR.sup.13--(CH.sub.2).sub.m(CHOR.sup.8).sub.n-
CH.sub.2NR.sup.13--(Z).sub.g-CAP,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-NH--C(.dbd.O)--NH--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m--C(.dbd.O)NR.-
sup.10R.sup.10,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.11R.sup.11,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.12R.sup.12,
Link-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--(Z).sub.g-CAP,
Link-(Z).sub.g--(CH.sub.2).sub.m-Het-(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CR.sup.11R.sup.11-CA-
P, Link
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2--CR.sup.11R.sup.11-CAP,
Link --(CH.sub.2).sub.n--(Z).sub.g--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n(Z).sub.g--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n--NR.sup.13--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.n--(CHOR.sup.8).sub.mCH.sub.2--NR.sup.13--(Z).sub.g---
CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.nNR.sup.13--(CH.sub.2).sub.m(CHOR.sup.8).sub.nCH.sub.-
2NR.sup.13--(Z).sub.g--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP-
, Link NH--C(.dbd.O)--NH--(CH.sub.2).sub.m--CR.sup.11R.sup.11-CAP,
Link-(CH.sub.2).sub.m--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m--CR.sup.11R.s-
up.11-CAP,
Link-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--(Z).sub.g---
CR.sup.11R.sup.11-CAP, or
Link-(Z).sub.g--(CH.sub.2).sub.m-Het-(CH.sub.2).sub.m--CR.sup.11R.sup.11--
CAP.
In a preferred embodiment, R.sup.5 is --OH,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8.
In another preferred embodiment R.sup.5 is one of the
following:
--(CH.sub.2).sub.m--OR.sup.8, --(CH.sub.2).sub.4--OH,
--O--(CH.sub.2).sub.m--OR.sup.8, --O--(CH.sub.2).sub.4--OH,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10, --NHSO.sub.2CH.sub.3,
--CH.sub.2NH(C.dbd.O)--(OCH.sub.3).sub.3, --NH(C.dbd.O)CH.sub.3,
--CH.sub.2NH.sub.2, --NH--CO.sub.2C.sub.2H.sub.5,
--CH.sub.2NH(C.dbd.O)CH.sub.3, --CH.sub.2NHCO.sub.2CH.sub.3,
--CH.sub.2NHSO.sub.2CH.sub.3,
--(CH.sub.2).sub.4--NH(C.dbd.O)O(CH.sub.3).sub.3,
--(CH.sub.2).sub.4--NH.sub.2,
--(CH.sub.2).sub.3--NH(C.dbd.O)O(CH.sub.3).sub.3,
--(CH.sub.2).sub.3--NH.sub.2,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--OCH.sub.2CH.sub.2NHCO.sub.2(CH.sub.3).sub.3,
--OCH.sub.2CH.sub.2NHCO.sub.2C.sub.2H.sub.5,
--O--(CH.sub.2).sub.3--NH--CO.sub.2--(CH.sub.3).sub.3,
--O(CH.sub.2).sub.3--NH.sub.2,
--OCH.sub.2CH.sub.2NHSO.sub.2CH.sub.3,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--OCH.sub.2CHOHCH.sub.2O-glucuronide,
--OCH.sub.2CH.sub.2CHOHCH.sub.2OH,
--OCH.sub.2-(.alpha.-CHOH).sub.2CH.sub.2OH,
--OCH.sub.2--(CHOH).sub.2CH.sub.2OH,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10, --C(.dbd.O)NH.sub.2,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--CH.sub.2--(C.dbd.O)NHCH.sub.2CHOH,
--O--CH.sub.2--(C.dbd.O)NHCH.sub.2CHOHCH.sub.2OH,
--O--CH.sub.2(C.dbd.O)NHCH.sub.2(CHOH).sub.2CH.sub.2OH,
--O--CH.sub.2C(C.dbd.O)NHSO.sub.2CH.sub.3,
--O--CH.sub.2(C.dbd.O)NHCO.sub.2CH.sub.3,
--O--CH.sub.2--C(C.dbd.O)NH--C(C.dbd.O)NH.sub.2,
--O--CH.sub.2--(C.dbd.O)NH--(C.dbd.O)CH.sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--(C.dbd.N)--NH.sub.2, --(C.dbd.NH)NH.sub.2,
--(CH.sub.2).sub.n--NH--C(.dbd.NH)--NH.sub.2,
--(CH.sub.2).sub.3--NH--C(.dbd.NH)--NH.sub.2,
--CH.sub.2NH--C(.dbd.NH)--NH.sub.2,
--(CH.sub.2).sub.n--CONHCH.sub.2(CHOH).sub.n--CH.sub.2OH,
--NH--C(.dbd.O)--CH.sub.2--(CHOH).sub.nCH.sub.2OH,
--NH--(C.dbd.O)--NH--CH.sub.2(CHOH).sub.2CHOH,
--NHC(C.dbd.O)NHCH.sub.2CH.sub.2OH,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--O(CH.sub.2).sub.3--NH--C(.dbd.NH)--NH.sub.2,
--O--(CH.sub.2).sub.m--CHNH.sub.2--CO.sub.2NR.sup.7R.sup.10,
--OCH.sub.2--CHNH.sub.2--CO.sub.2NH.sub.2,
--O--(CH.sub.2).sub.m--CHNH.sub.2--CO.sub.2NR.sup.7R.sup.10
(anomeric center is the (R) enantiomer),
--O--(CH.sub.2).sub.m--CHNH.sub.2--CO.sub.2NR.sup.7R.sup.10
(anomeric center is the (S) enantiomer),
--OCH.sub.2CHOH--CH.sub.2NHCO.sub.2(CH.sub.3).sub.3,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8, --NHCH.sub.2(CHOH).sub.2CH.sub.2OH,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --O--(CH.sub.2).sub.m--CO.sub.2R.sup.7,
--OCH.sub.2CH.sub.2CO.sub.2(CH.sub.3).sub.3, --OCH.sub.2CO.sub.2H,
--OCH.sub.2CO.sub.2C.sub.2H.sub.5, --O--(CH.sub.2).sub.m-Boc,
--(CH.sub.2).sub.m-Boc,
--O--(CH.sub.2).sub.m--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.n--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.m--NH--C(.dbd.O)--OR.sup.7,
--O--(CH.sub.2).sub.m--NH--C(.dbd.O)--OR.sup.7,
--(CH.sub.2).sub.n--NH--C(.dbd.O)--R.sup.11,
--O--(CH.sub.2).sub.m--NH--C(.dbd.O)--R.sup.11,
--O--(CH.sub.2).sub.m--C(.dbd.O)N(R.sup.7).sub.2,
--(CH.sub.2).sub.m--CHOH--CH.sub.2--NHBoc,
--O--(CH.sub.2).sub.m--CHOH--CH.sub.2--NHBoc,
--(CH.sub.2).sub.m--NHC(O)OR.sup.7,
--O--(CH.sub.2).sub.m--NHC(O)OR.sup.7,
--O--(CH.sub.2).sub.m--C(.dbd.NH)--N(R.sup.7).sub.2, or
--(CH.sub.2).sub.n--C(.dbd.NH)--N(R.sup.7).sub.2.
In another embodiment, R.sup.5 is selected from the group
consisting of --O--(CH.sub.2).sub.3--OH, --NH.sub.2,
--O--CH.sub.2--(CHOH).sub.2--CH.sub.2OH--O--CH.sub.2--CHOH--CH.sub.2OH,
--O--CH.sub.2CH.sub.2--O-tetrahydropyran-2-yl,
--O--CH.sub.2CHOH--CH.sub.2--O-glucuronide,
--O--CH.sub.2CH.sub.2OH, --O--(CH.sub.2CH.sub.2O).sub.4--CH.sub.3,
--O--CH.sub.2CH.sub.2OCH.sub.3,
--O--CH.sub.2--(CHOC(.dbd.O)CH.sub.3)--CH.sub.2--OC(.dbd.O)CH.sub.3,
--O--(CH.sub.2CH.sub.2O).sub.2--CH.sub.3,
--OCH.sub.2--CHOH--CHOH--CH.sub.2OH, --CH.sub.2OH,
--CO.sub.2CH.sub.3,
##STR00023##
In another embodiment, R.sup.5 is selected from the group
consisting of --O--(CH.sub.2).sub.3--OH, --NH.sub.2,
--O--CH.sub.2--(CHOH).sub.2--CH.sub.2OH,
--O--CH.sub.2--CHOH--CH.sub.2OH,
--O--CH.sub.2CH.sub.2--O-tetrahydropyran-2-yl,
--O--CH.sub.2CHOH--CH.sub.2--O-glucuronide,
--O--CH.sub.2CH.sub.2OH, --O--(CH.sub.2CH.sub.2O).sub.4--CH.sub.3,
--O--CH.sub.2CH.sub.2OCH.sub.3,
--O--CH.sub.2--(CHOC(.dbd.O)CH.sub.3)--CH.sub.2--OC(.dbd.O)CH.sub.3,
--O--(CH.sub.2CH.sub.2O).sub.2--CH.sub.3,
--OCH.sub.2--CHOH--CHOH--CH.sub.2OH, --CH.sub.2OH,
--CO.sub.2CH.sub.3, --SO.sub.3H, --O-glucuronide,
##STR00024##
In a preferred embodiment, each
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7 falls within the scope of
the structures described above and is, independently,
--(CH.sub.2).sub.n--(C.dbd.N)--NH.sub.2,
--(CH.sub.2).sub.n--NH--C(.dbd.NH)NH.sub.2,
--(CH.sub.2).sub.n--CONHCH.sub.2(CHOH).sub.n--CH.sub.2OH, or
--NH--C(.dbd.O)--CH.sub.2--(CHOH).sub.nCH.sub.2OH.
In another a preferred embodiment, each
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7 falls within the scope of
the structures described above and is, independently,
--O--(CH.sub.2).sub.m--NH--C(.dbd.NH)--N(R.sup.7).sub.2, or
--O--(CH.sub.2).sub.m--CHNH.sub.2--CO.sub.2NR.sup.7R.sup.10.
In another preferred embodiment, R.sup.5 may be one of the
following: --O--CH.sub.2CHOHCH.sub.2O-glucuronide,
--OCH.sub.2CHOHCH.sub.3, --OCH.sub.2CH.sub.2NH.sub.2,
--OCH.sub.2CH.sub.2NHCO(CH.sub.3).sub.3, --CH.sub.2CH.sub.2OH,
--OCH.sub.2CH.sub.2OH, --O--(CH.sub.2).sub.m-Boc,
--(CH.sub.2).sub.m-Boc, --OCH.sub.2CH.sub.2OH,
--OCH.sub.2CO.sub.2H,
--O--(CH.sub.2).sub.m--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.n--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--NHCH.sub.2(CHOH).sub.2--CH.sub.2OH, --OCH.sub.2CO.sub.2Et,
--NHSO.sub.2CH.sub.3, --(CH.sub.2).sub.m--NH--C(.dbd.O)--OR.sup.7,
--O--(CH.sub.2).sub.m--NH--C(.dbd.O)--OR.sup.7,
--(CH.sub.2).sub.n--NH--C(.dbd.O)--R.sup.11,
--O--(CH.sub.2).sub.m--NH--C(.dbd.O)--R.sup.11,
--O--CH.sub.2C(.dbd.O)NH.sub.2, --CH.sub.2NH.sub.2, --NHCO.sub.2Et,
--OCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH,
--CH.sub.2NHSO.sub.2CH.sub.3, --OCH.sub.2CH.sub.2CHOHCH.sub.2OH,
--OCH.sub.2CH.sub.2NHCO.sub.2Et, --NH--C(.dbd.NH2)-NH.sub.2,
--OCH.sub.2-(.alpha.-CHOH).sub.2--CH.sub.2OH,
--OCH.sub.2CHOHCH.sub.2NH.sub.2,
--(CH.sub.2).sub.m--CHOH--CH.sub.2--NHBoc,
--O--(CH.sub.2).sub.m--CHOH--CH.sub.2--NHBoc,
--(CH.sub.2).sub.m--NHC(O)OR.sup.7,
--O--(CH.sub.2).sub.m--NHC(O)OR.sup.7,
--OCH.sub.2CH.sub.2CH.sub.2NH.sub.2,
--OCH.sub.2CH.sub.2NHCH.sub.2(CHOH).sub.2CH.sub.2OH,
--OCH.sub.2CH.sub.2NH(CH.sub.2[(CHOH).sub.2CH.sub.2OH)].sub.2,
--(CH.sub.2).sub.4--NHBoc, --(CH.sub.2).sub.4--NH.sub.2,
--(CH.sub.2).sub.4--OH, --OCH.sub.2CH.sub.2NHSO.sub.2CH.sub.3,
--O--(CH.sub.2).sub.m--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.n--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.3--NH Boc, --(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.m--NH--NH--C(.dbd.NH)--N(R.sup.7).sub.2,
--(CH.sub.2).sub.n--NH--NH--C(.dbd.NH)--N(R.sup.7).sub.2, or
--O--CH.sub.2--CHOH--CH.sub.2--NH--C(.dbd.NH)--N(R.sup.7).sub.2.
In another preferred embodiment, R.sup.5 is --OH,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8.
In a particularly preferred embodiment, R.sup.5 is
--O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00025##
Selected substituents within the compounds of the invention are
present to a recursive degree. In this context, "recursive
substituent" means that a substituent may recite another instance
of itself. Because of the recursive nature of such substituents,
theoretically, a large number of compounds may be present in any
given embodiment. For example, R.sup.9 contains a R.sup.13
substituent. R.sup.13 can contain an R.sup.10 substituent and
R.sup.10 can contain a R.sup.9 substituent. One of ordinary skill
in the art of medicinal chemistry understands that the total number
of such substituents is reasonably limited by the desired
properties of the compound intended. Such properties include, by
way of example and not limitation, physical properties such as
molecular weight, solubility or log P, application properties such
as activity against the intended target, and practical properties
such as ease of synthesis.
By way of example and not limitation, R.sup.9, R.sup.13 and
R.sup.10 are recursive substituents in certain embodiments.
Typically, each of these may independently occur 20, 19, 18, 17,
16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0, times
in a given embodiment. More typically, each of these may
independently occur 12 or fewer times in a given embodiment. More
typically yet, R.sup.9 will occur 0 to 8 times in a given
embodiment, R.sup.13 will occur 0 to 6 times in a given embodiment
and R.sup.10 will occur 0 to 6 times in a given embodiment. Even
more typically yet, R.sup.9 will occur 0 to 6 times in a given
embodiment, R.sup.13 will occur 0 to 4 times in a given embodiment
and R.sup.10 will occur 0 to 4 times in a given embodiment.
Recursive substituents are an intended aspect of the invention. One
of ordinary skill in the art of medicinal chemistry understands the
versatility of such substituents. To the degree that recursive
substituents are present in an embodiment of the invention, the
total number will be determined as set forth above.
Each -Het- is, independently, --N(R.sup.7)--, --N(R.sup.10)--,
--S--, --SO--, --SO.sub.2--; --O--, --SO.sub.2NH--, --NHSO.sub.2--,
--NR.sup.7CO--, --CONR.sup.7--, --N(R.sup.13)--,
--SO.sub.2NR.sup.13--, --NR.sup.13CO--, or --CONR.sup.13--. In a
preferred embodiment, -Het- is --O--, --N(R.sup.7)--, or
--N(R.sup.10)--. Most preferably, -Het- is --O--.
Each -Link- is, independently, --O--, --(CH.sub.2).sub.n--,
--O(CH.sub.2).sub.m--, --NR.sup.13--C(.dbd.O)--NR.sup.13--,
--NR.sup.13--C(.dbd.O)--(CH.sub.2).sub.m--,
--C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m.sup.-,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.n.sup.-, --S--,
--SO--, --SO.sub.2--, --SO.sub.2NR.sup.7--, --SO.sub.2NR.sup.10--,
or -Het-. In a preferred embodiment, -Link- is --O--,
--(CH.sub.2).sub.n--, --NR.sup.13--C(.dbd.O)--(CH.sub.2).sub.m--,
or --C(.dbd.O)NR.sup.13--(CH.sub.2).sub.m.sup.-.
Each -CAP is, independently, thiazolidinedione, oxazolidinedione,
-heteroaryl-C(.dbd.O)NR.sup.13R.sup.13, heteroaryl-W, --CN,
--O--C(.dbd.S)NR.sup.13R.sup.13, --(Z).sub.gR.sup.13,
--CR.sup.10((Z).sub.gR.sup.13)((Z).sub.gR.sup.13), --C(.dbd.O)OAr,
--C(.dbd.O)NR.sup.13Ar, imidazoline, tetrazole, tetrazole amide,
--SO.sub.2NHR.sup.13,
--SO.sub.2NH--C(R.sup.13R.sup.13)--(Z).sub.g--R.sup.13, a cyclic
sugar or oligosaccharide, a cyclic amino sugar, oligosaccharide,
--CR.sup.10
(--(CH.sub.2).sub.m--R.sup.9)(--(CH.sub.2).sub.m--R.sup.9),
--N(--(CH.sub.2).sub.m--R.sup.9)(--(CH.sub.2).sub.m--R.sup.9),
--NR.sup.13(--(CH.sub.2).sub.m--CO.sub.2R.sup.13),
##STR00026##
In a preferred embodiment, CAP is
##STR00027##
Each Ar is, independently, phenyl, substituted phenyl, wherein the
substituents of the substituted phenyl are 1-3 substituents
independently selected from the group consisting of OH, OCH.sub.3,
NR.sup.13R.sup.13, Cl, F, and CH.sub.3, or heteroaryl.
Examples of heteroaryl include pyridinyl, pyrazinyl, furanyl,
thienyl, tetrazolyl, thiazolidinedionyl, imidazoyl, pyrrolyl,
quinolinyl, indolyl, adeninyl, pyrazolyl, thiazolyl, isoxazolyl,
benzimidazolyl, purinyl, isoquinolinyl, pyridazinyl, pyrimidinyl,
1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, cinnolinyl,
phthalazinyl, quinazolinyl, quinoxalinyl, and pterdinyl groups.
Each W is, independently, thiazolidinedione, oxazolidinedione,
heteroaryl-C(.dbd.O)N R.sup.13R.sup.13, --CN,
--O--C(.dbd.S)NR.sup.13R.sup.13, --(Z).sub.gR.sup.13,
--CR.sup.10((Z).sub.gR.sup.13)((Z).sub.gR.sup.13), --C(.dbd.O)OAr,
--C(.dbd.O)N R.sup.13Ar, imidazoline, tetrazole, tetrazole amide,
--SO.sub.2NHR.sup.13,
--SO.sub.2NH--C(R.sup.13R.sup.13)--(Z).sub.g--R.sup.13, a cyclic
sugar or oligosaccharide, a cyclic amino sugar,
oligosaccharide,
##STR00028##
There is at least one R.sup.5 on A.sup.1 and A.sup.2 and the
remaining substituents are R.sup.6. Each R.sup.6 is, independently,
R.sup.5, --R.sup.7, --OR.sup.11, --N(R.sup.7).sub.2,
--(CH.sub.2).sub.m--OR.sup.8, --O--(CH.sub.2).sub.m--OR.sup.8,
--(CH.sub.2).sub.n--NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--NR.sup.7R.sup.10,
--(CH.sub.2).sub.n(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--O--(CH.sub.2CH.sub.2O).sub.m--R.sup.8,
--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--O--(CH.sub.2CH.sub.2O).sub.m--CH.sub.2CH.sub.2NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--(CH.sub.2).sub.n--(Z).sub.g--R.sup.7,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7,
--(CH.sub.2).sub.n--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--CH-
.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8, --(CH.sub.2).sub.n--CO.sub.2R.sup.7,
--O--(CH.sub.2).sub.m--CO.sub.2R.sup.7, --OSO.sub.3H,
--O-glucuronide, --O-glucose,
##STR00029##
When two R.sup.6 are --OR.sup.11 and are located adjacent to each
other on the aromatic carbocycle or aromatic heterocycle, the two
OR.sup.11 may form a methylenedioxy group; i.e., a group of the
formula --O--CH.sub.2--O--.
In addition, one or more of the R.sup.6 groups can be one of the
R.sup.5 groups which fall within the broad definition of R.sup.6
set forth above.
R.sup.6 may be hydrogen. Therefore, provided that the aromatic
carbocycle or aromatic heterocycle is substituted with R.sup.5, the
remaining R.sup.6 may be hydrogen. Preferably, at most, 3 of the
R.sup.6 groups are other than hydrogen. More preferably, provided
that the aromatic carbocycle or aromatic heterocycle is substituted
with R.sup.5, then R.sup.6 is H.
Each g is, independently, an integer from 1 to 6. Therefore, each g
may be 1, 2, 3, 4, 5, or 6.
Each m is an integer from 1 to 7. Therefore, each m may be 1, 2, 3,
4, 5, 6, or 7.
Each n is an integer from 0 to 7. Therefore, each n may be 0, 1, 2,
3, 4, 5, 6, or 7.
Each Z is, independently, --(CHOH)--, --C(.dbd.O)--,
--(CHNR.sup.7R.sup.10)--, --(C.dbd.NR.sup.10)--, --NR.sup.10--,
--(CH.sub.2).sub.n--, --(CHNR.sup.13R.sup.13)--,
--(C.dbd.NR.sup.13)--, or --NR.sup.13--. As designated by (Z).sub.g
in certain embodiments, Z may occur one, two, three, four, five or
six times and each occurance of Z is, independently, --(CHOH)--,
--C(.dbd.O)--, --(CHNR.sup.7R.sup.10)--, --(C.dbd.NR.sup.10)--,
--NR.sup.10--, --(CH.sub.2).sub.n--, --(CHNR.sup.13R.sup.13)--,
--(C.dbd.NR.sup.13)--, or --NR.sup.13--. Therefore, by way of
example and not by way of limitation, (Z).sub.g can be
--(CHOH)--(CHNR.sup.7R.sup.10)--,
--(CHOH)--(CHNR.sup.7R.sup.10)--C(.dbd.O)--,
--(CHOH)--(CHNR.sup.7R.sup.10)--C(.dbd.O)--(CH.sub.2).sub.n--,
--(CHOH)--(CHNR.sup.7R.sup.10)--C(.dbd.O)--(CH.sub.2).sub.n--(CHNR.sup.13-
R.sup.13)--,
--(CHOH)--(CHNR.sup.7R.sup.10)--C(.dbd.O)--(CH.sub.2).sub.n--(CHNR.sup.13-
R.sup.13)--C(.dbd.O)--, and the like.
In any variable containing --CHOR.sup.8-- or --CH.sub.2OR.sup.8
groups, when any --CHOR.sup.8-- or --CH.sub.2OR.sup.8 groups are
located 1,2- or 1,3- with respect to each other, the R.sup.8 groups
may, optionally, be taken together to form a cyclic mono- or
di-substituted 1,3-dioxane or 1,3-dioxolane.
More specific examples of suitable compounds represented by formula
(I) are shown in formulas II and III below wherein A.sup.1 and
A.sup.2 are defined as above:
##STR00030##
In a preferred aspect of formula II, A.sup.1 is selected from
indenyl, napthalenyl, 1,2-dihydronapthalenyl,
1,2,3,4-tetrahydronapthalenyl, anthracenyl, fluorenyl,
phenanthrenyl, azulenyl, cyclohepta-1,3,5-trienyl or
5H-dibenzo[a,d]cycloheptenyl.
In another preferred aspect of formula II, A.sup.1 is
##STR00031##
wherein each Q is, independently, C--H, C--R.sup.5, or C--R.sup.6,
with the proviso that at least one Q is C--R.sup.5. Preferably, six
Q are C--H. Preferably, each R.sup.6 is H. Preferably, R.sup.5 is
--OH, --O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.mN.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8. More preferably, six Q are C--H and R.sup.5 is
--OH, --O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--,
(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8. More preferably, R.sup.5 is
--O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00032##
Most preferably, R.sup.5--O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00033## and six Q are C--H.
In another preferred aspect of formula II, A.sup.1 is
##STR00034##
Preferably, R.sup.5 is --OH,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8. Most preferably, R.sup.5 is
--O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00035##
In a preferred aspect of formula III, A.sup.2 is selected from
indolizinyl, indolyl, isoindolyl, indolinyl, benzo[b]furanyl,
2,3-dihydrobenzo[b]furanyl, benzo[b]thiophenyl,
2,3-dihydrobenzo[b]thiophenyl, indazolyl, benzimidazolyl,
benzthiazolyl, purinyl, quinolinyl, 1,2,3,4-tetrahydroquinolinyl,
3,4-dihydro-2H-chromenyl, 3,4-dihydro-2H-thiochromenyl,
isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl,
quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl,
acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl,
dibenzofuranyl, dibenzothiophenyl, 1H-azepinyl,
5H-dibenz[b,f]azepinyl, or
10,11-dihydro-5H-dibenz[b,f]azepinyl.
In another preferred aspect of formula III, A.sup.2 is
##STR00036## wherein each Q is, independently, C--H, C--R.sup.5,
C--R.sup.6, or a nitrogen atom, with the proviso that at least one
Q is nitrogen and one Q is C--R.sup.5, and at most three Q in a
ring are nitrogen atoms. In a preferred embodiment, only one Q in
each ring is nitrogen. In another preferred embodiment, only a
single Q is nitrogen. In a particularly preferred embodiment, a
single Q is nitrogen, one Q is C--R.sup.5, and the remaining Q are
C--H. In another preferred embodiment, each R.sup.6 is H.
Preferably, R.sup.5 is --OH,
--O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m--(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8. More preferably, one Q is nitrogen, five Q are
C--H and R.sup.5 is --OH, --O--(CH.sub.2).sub.m(Z).sub.gR.sup.12,
-Het-(CH.sub.2).sub.m--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.13,
-Het-(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.mNH--C(.dbd.NR.sup.13)---
NR.sup.13R.sup.13,
-Link-(CH.sub.2).sub.m--(Z).sub.g--(CH.sub.2).sub.m-CAP,
Link-(CH.sub.2).sub.n--CR.sup.11R.sup.11-CAP,
-Het-(CH.sub.2).sub.m--CONR.sup.13R.sup.13,
--(CH.sub.2).sub.n--NR.sup.12R.sup.12,
--O--(CH.sub.2).sub.mNR.sup.11R.sup.11,
--O--(CH.sub.2).sub.m--N.sup..sym.--(R.sup.11).sub.3,
--(CH.sub.2).sub.n--(Z).sub.g--(CH.sub.2).sub.m--NR.sup.10R.sup.10,
-Het-(CH.sub.2).sub.m--(Z).sub.g--NH--C(.dbd.NR.sup.13)--NR.sup.13R.sup.1-
3,
--O--(CH.sub.2).sub.m(CHOR.sup.8)(CHOR.sup.8).sub.n--CH.sub.2OR.sup.8,
--O--(CH.sub.2).sub.m--C(.dbd.O)NR.sup.7R.sup.10,
--O--(CH.sub.2).sub.m(Z).sub.g--R.sup.7, or
--O--(CH.sub.2).sub.m--NR.sup.10--CH.sub.2(CHOR.sup.8)(CHOR.sup.8).sub.n--
-CH.sub.2OR.sup.8. More preferably, R.sup.5 is
--O--CH.sub.2--(CHOH)--CH.sub.2OH, --OH,
--O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00037##
Most preferably, R.sup.5 is --O--CH.sub.2--(CHOH)--CH.sub.2OH,
--OH, --O--(CH.sub.2).sub.3NH.sub.2,
--O--(CH.sub.2).sub.3NH(C.dbd.NH)NH.sub.2,
--O--(CH.sub.2).sub.2NH(C.dbd.NH)NH.sub.2,
--O--CH.sub.2(CO)NH.sub.2,
--O--(CH.sub.2).sub.2--N.sup..sym.--(CH.sub.3).sub.3,
##STR00038## a single Q is nitrogen and five Q are C--H.
In a particularly preferred embodiment, the compounds of formula I,
formula II, or formula III are:
##STR00039##
In another preferred embodiment, the compounds of the present
invention are represented by the following formulas:
##STR00040##
The compounds described herein may be prepared and used as the free
base. Alternatively, the compounds may be prepared and used as a
pharmaceutically acceptable salt. Pharmaceutically acceptable salts
are salts that retain or enhance the desired biological activity of
the parent compound and do not impart undesired toxicological
effects. Examples of such salts are (a) acid addition salts formed
with inorganic acids, for example, hydrochloric acid, hydrobromic
acid, sulfuric acid, phosphoric acid, nitric acid and the like; (b)
salts formed with organic acids such as, for example, acetic acid,
oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric
acid, gluconic acid, citric acid, malic acid, ascorbic acid,
benzoic acid, tannic acid, palmitic acid, alginic acid,
polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid,
p-toluenesulfonic acid, naphthalenedisulfonic acid,
polygalacturonic acid, malonic acid, sulfosalicylic acid, glycolic
acid, 2-hydroxy-3-naphthoate, pamoate, salicylic acid, stearic
acid, phthalic acid, mandelic acid, lactic acid and the like; and
(c) salts formed from elemental anions for example, chlorine,
bromine, and iodine.
It is to be noted that all enantiomers, diastereomers, and racemic
mixtures, tautomers, polymorphs, pseudopolymorphs and
pharmaceutically acceptable salts of compounds within the scope of
formula (I), formula II, or formula III are embraced by the present
invention. All mixtures of such enantiomers and diastereomers are
within the scope of the present invention.
A compound of formula I-III and its pharmaceutically acceptable
salts may exist as different polymorphs or pseudopolymorphs. As
used herein, crystalline polymorphism means the ability of a
crystalline compound to exist in different crystal structures. The
crystalline polymorphism may result from differences in crystal
packing (packing polymorphism) or differences in packing between
different conformers of the same molecule (conformational
polymorphism). As used herein, crystalline pseudopolymorphism means
the ability of a hydrate or solvate of a compound to exist in
different crystal structures. The pseudopolymorphs of the instant
invention may exist due to differences in crystal packing (packing
pseudopolymorphism) or due to differences in packing between
different conformers of the same molecule (conformational
pseudopolymorphism). The instant invention comprises all polymorphs
and pseudopolymorphs of the compounds of formula I-III and their
pharmaceutically acceptable salts.
A compound of formula I-III and its pharmaceutically acceptable
salts may also exist as an amorphous solid. As used herein, an
amorphous solid is a solid in which there is no long-range order of
the positions of the atoms in the solid. This definition applies as
well when the crystal size is two nanometers or less. Additives,
including solvents, may be used to create the amorphous forms of
the instant invention. The instant invention comprises all
amorphous forms of the compounds of formula I-III and their
pharmaceutically acceptable salts.
The compounds of formula I-III may exist in different tautomeric
forms. One skilled in the art will recognize that amidines, amides,
guanidines, ureas, thioureas, heterocycles and the like can exist
in tautomeric forms. By way of example and not by way of
limitation, compounds of formula I-III can exist in various
tautomeric forms as shown below:
##STR00041## All possible tautomeric forms of the amidines, amides,
guanidines, ureas, thioureas, heterocycles and the like of all of
the embodiments of formula I-III are within the scope of the
instant invention.
"Enantiomers" refer to two stereoisomers of a compound which are
non-superimposable mirror images of one another.
Stereochemical definitions and conventions used herein generally
follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms
(1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen,
S., Stereochemistry of Organic Compounds (1994) John Wiley &
Sons, Inc., New York. Many organic compounds exist in optically
active forms, i.e., they have the ability to rotate the plane of
plane-polarized light. In describing an optically active compound,
the prefixes D and L or R and S are used to denote the absolute
configuration of the molecule about its chiral center(s). The
prefixes d and l, D and L, or (+) and (-) are employed to designate
the sign of rotation of plane-polarized light by the compound, with
S, (-), or l meaning that the compound is levorotatory while a
compound prefixed with R, (+), or d is dextrorotatory. For a given
chemical structure, these stereoisomers are identical except that
they are mirror images of one another. A specific stereoisomer may
also be referred to as an enantiomer, and a mixture of such isomers
is often called an enantiomeric mixture. A 50:50 mixture of
enantiomers is referred to as a racemic mixture or a racemate,
which may occur where there has been no stereoselection or
stereospecificity in a chemical reaction or process. The terms
"racemic mixture" and "racemate" refer to an equimolar mixture of
two enantiomeric species, devoid of optical activity.
A single stereoisomer, e.g. an enantiomer, substantially free of
its stereoisomer may be obtained by resolution of the racemic
mixture using a method such as formation of diastereomers using
optically active resolving agents ("Stereochemistry of Carbon
Compounds," (1962) by E. L. Eliel, McGraw Hill; Lochmuller, C. H.,
(1975) J. Chromatogr., 113:(3) 283-302). Racemic mixtures of chiral
compounds of the invention can be separated and isolated by any
suitable method, including: (1) formation of ionic, diastereomeric
salts with chiral compounds and separation by fractional
crystallization or other methods, (2) formation of diastereomeric
compounds with chiral derivatizing reagents, separation of the
diastereomers, and conversion to the pure stereoisomers, and (3)
separation of the substantially pure or enriched stereoisomers
directly under chiral conditions.
"Diastereomer" refers to a stereoisomer with two or more centers of
chirality and whose molecules are not mirror images of one another.
Diastereomers have different physical properties, e.g. melting
points, boiling points, spectral properties, and reactivities.
Mixtures of diastereomers may separate under high resolution
analytical procedures such as electrophoresis and
chromatography.
Without being limited to any particular theory, it is believed that
the compounds of formula (I), formula II, or formula III function
in vivo as sodium channel blockers. By blocking epithelial sodium
channels present in mucosal surfaces the compounds of formula (I),
formula II, or formula III reduce the absorption of water by the
mucosal surfaces. This effect increases the volume of protective
liquids on mucosal surfaces, rebalances the system, and thus treats
disease.
The present invention also provides methods of treatment that take
advantage of the properties of the compounds described herein as
discussed above. Thus, subjects that may be treated by the methods
of the present invention include, but are not limited to, patients
afflicted with cystic fibrosis, primary ciliary dyskinesia, chronic
bronchitis, bronchiectasis chronic obstructive airway disease,
artificially ventilated patients, patients with acute pneumonia,
etc. The present invention may be used to obtain a sputum sample
from a patient by administering the active compounds to at least
one lung of a patient, and then inducing or collecting a sputum
sample from that patient. Typically, the invention will be
administered to respiratory mucosal surfaces via aerosol (liquid or
dry powders) or lavage.
Subjects that may be treated by the method of the present invention
also include patients being administered supplemental oxygen
nasally (a regimen that tends to dry the airway surfaces); patients
afflicted with an allergic disease or response (e.g., an allergic
response to pollen, dust, animal hair or particles, insects or
insect particles, etc.) that affects nasal airway surfaces;
patients afflicted with a bacterial infection e.g., staphylococcus
infections such as Staphylococcus aureus infections, Hemophilus
influenza infections, Streptococcus pneumoniae infections,
Pseudomonas aeuriginosa infections, etc.) of the nasal airway
surfaces; patients afflicted with an inflammatory disease that
affects nasal airway surfaces; or patients afflicted with sinusitis
(wherein the active agent or agents are administered to promote
drainage of congested mucous secretions in the sinuses by
administering an amount effective to promote drainage of congested
fluid in the sinuses), or combined, Rhinosinusitis. The invention
may be administered to rhino-sinal surfaces by topical delivery,
including aerosols and drops.
The present invention may be used to hydrate mucosal surfaces other
than airway surfaces. Such other mucosal surfaces include
gastrointestinal surfaces, oral surfaces, genito-urethral surfaces,
ocular surfaces or surfaces of the eye, the inner ear and the
middle ear. For example, the active compounds of the present
invention may be administered by any suitable means, including
locally/topically, orally, or rectally, in an effective amount.
The compounds of the present invention are also useful for treating
a variety of functions relating to the cardiovascular system. Thus,
the compounds of the present invention are useful for use as
antihypertensive agents. The compounds may also be used to reduce
blood pressure and to treat edema. In addition, the compounds of
the present invention are also useful for promoting diuresis,
natriuresis, and saluresis. The compounds may be used alone or in
combination with beta blockers, ACE inhibitors, HMGCoA reductase
inhibitors, calcium channel blockers and other cardiovascular
agents to treat hypertension, congestive heart failure and reduce
cardiovascular mortality.
The present invention is concerned primarily with the treatment of
human subjects, but may also be employed for the treatment of other
mammalian subjects, such as dogs and cats, for veterinary
purposes.
As discussed above, the compounds used to prepare the compositions
of the present invention may be in the form of a pharmaceutically
acceptable free base. Because the free base of the compound is
generally less soluble in aqueous solutions than the salt, free
base compositions are employed to provide more sustained release of
active agent to the lungs. An active agent present in the lungs in
particulate form which has not dissolved into solution is not
available to induce a physiological response, but serves as a depot
of bioavailable drug which gradually dissolves into solution.
Another aspect of the present invention is a pharmaceutical
composition, comprising a compound of formula (I), formula II, or
formula III in a pharmaceutically acceptable carrier (e.g., an
aqueous carrier solution). In general, the compound of formula (I),
formula II, or formula III is included in the composition in an
amount effective to inhibit the reabsorption of water by mucosal
surfaces.
Without being limited to any particular theory, it is believed that
sodium channel blockers of the present invention block epithelial
sodium channels present in mucosal surfaces the sodium channel
blocker, described herein reduce the absorption of salt and water
by the mucosal surfaces. This effect increases the volume of
protective liquids on mucosal surfaces, rebalances the system, and
thus treats disease. This effect is enhanced when used in
combination with osmolytes.
The compounds of formula (I), formula II, or formula III may also
be used in conjunction with osmolytes thus lowering the dose of the
compound needed to hydrate mucosal surfaces. This important
property means that the compound will have a lower tendency to
cause undesired side-effects by blocking sodium channels located at
untargeted locations in the body of the recipient, e.g., in the
kidneys when used in combination with an osmolyte.
Active osmolytes of the present invention are molecules or
compounds that are osmotically active (i.e., are "osmolytes").
"Osmotically active" compounds of the present invention are
membrane-impermeable (i.e., essentially non-absorbable) on the
airway or pulmonary epithelial surface. The terms "airway surface"
and "pulmonary surface," as used herein, include pulmonary airway
surfaces such as the bronchi and bronchioles, alveolar surfaces,
and nasal and sinus surfaces. Active compounds of the present
invention may be ionic osmolytes (i.e., salts), or may be non-ionic
osmolytes (i.e., sugars, sugar alcohols, and organic osmolytes). It
is specifically intended that both racemic forms of the active
compounds that are racemic in nature are included in the group of
active compounds that are useful in the present invention. It is to
be noted that all racemates, enantiomers, diastereomers, tautomers,
polymorphs and pseudopolymorphs and racemic mixtures of the
osmotically active compounds are embraced by the present
invention.
Active osmolytes useful in the present invention that are ionic
osmolytes include any salt of a pharmaceutically acceptable anion
and a pharmaceutically acceptable cation. Preferably, either (or
both) of the anion and cation are non-absorbable (i.e., osmotically
active and not subject to rapid active transport) in relation to
the airway surfaces to which they are administered. Such compounds
include but are not limited to anions and cations that are
contained in FDA approved commercially marketed salts, see, e.g.,
Remington: The Science and Practice of Pharmacy, Vol. II, pg. 1457
(19.sup.th Ed. 1995), incorporated herein by reference, and can be
used in any combination including their conventional
combinations.
Pharmaceutically acceptable osmotically active anions that can be
used to carry out the present invention include, but are not
limited to, acetate, benzenesulfonate, benzoate, bicarbonate,
bitartrate, bromide, calcium edetate, camsylate (camphorsulfonate),
carbonate, chloride, citrate, dihydrochloride, edetate, edisylate
(1,2-ethanedisulfonate), estolate (lauryl sulfate), esylate
(1,2-ethanedisulfonate), fumarate, gluceptate, gluconate,
glutamate, glycollylarsanilate (p-glycollamidophenylarsonate),
hexylresorcinate, hydrabamine
(N,N'-Di(dehydroabietyl)ethylenediamine), hydrobromide,
hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate,
lactobionate, malate, maleate, mandelate, mesylate, methylbromide,
methylnitrate, methylsulfate, mucate, napsylate, nitrate, nitrte,
pamoate (embonate), pantothenate, phosphate or diphosphate,
polygalacturonate, salicylate, stearate, subacetate, succinate,
sulfate, tannate, tartrate, teoclate (8-chlorotheophyllinate),
triethiodide, bicarbonate, etc. Particularly preferred anions
include chloride, sulfate, nitrate, gluconate, iodide, bicarbonate,
bromide, and phosphate.
Pharmaceutically acceptable cations that can be used to carry out
the present invention include, but are not limited to, organic
cations such as benzathine (N,N'-dibenzylethylenediamine),
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine
(N-methyl D-glucamine), procaine, D-lysine, L-lysine, D-arginine,
L-arginine, triethylammonium, N-methyl D-glycerol, and the like.
Particularly preferred organic cations are 3-carbon, 4-carbon,
5-carbon and 6-carbon organic cations. Metallic cations useful in
the practice of the present invention include but are not limited
to aluminum, calcium, lithium, magnesium, potassium, sodium, zinc,
iron, ammonium, and the like. Particularly preferred cations
include sodium, potassium, choline, lithium, meglumine, D-lysine,
ammonium, magnesium, and calcium.
Specific examples of osmotically active salts that may be used with
the sodium channel blockers described herein to carry out the
present invention include, but are not limited to, sodium chloride,
potassium chloride, choline chloride, choline iodide, lithium
chloride, meglumine chloride, L-lysine chloride, D-lysine chloride,
ammonium chloride, potassium sulfate, potassium nitrate, potassium
gluconate, potassium iodide, ferric chloride, ferrous chloride,
potassium bromide, etc. Either a single salt or a combination of
different osmotically active salts may be used to carry out the
present invention. Combinations of different salts are preferred.
When different salts are used, one of the anion or cation may be
the same among the differing salts.
Osmotically active compounds of the present invention also include
non-ionic osmolytes such as sugars, sugar-alcohols, and organic
osmolytes. Sugars and sugar-alcohols useful in the practice of the
present invention include but are not limited to 3-carbon sugars
(e.g., glycerol, dihydroxyacetone); 4-carbon sugars (e.g., both the
D and L forms of erythrose, threose, and erythrulose); 5-carbon
sugars (e.g., both the D and L forms of ribose, arabinose, xylose,
lyxose, psicose, fructose, sorbose, and tagatose); and 6-carbon
sugars (e.g., both the D and L forms of altose, allose, glucose,
mannose, gulose, idose, galactose, and talose, and the D and L
forms of allo-heptulose, allo-hepulose, gluco-heptulose,
manno-heptulose, gulo-heptulose, ido-heptulose, galacto-heptulose,
talo-heptulose). Additional sugars useful in the practice of the
present invention include raffinose, raffinose series
oligosaccharides, and stachyose. Both the D and L forms of the
reduced form of each sugar/sugar alcohol useful in the present
invention are also active compounds within the scope of the
invention. For example, glucose, when reduced, becomes sorbitol;
within the scope of the invention, sorbitol and other reduced forms
of sugar/sugar alcohols (e.g., mannitol, dulcitol, arabitol) are
accordingly active compounds of the present invention.
Osmotically active compounds of the present invention additionally
include the family of non-ionic osmolytes termed "organic
osmolytes." The term "organic osmolytes" is generally used to refer
to molecules used to control intracellular osmolality in the
kidney. See e.g., J. S. Handler et al., Comp. Biochem. Physiol,
117, 301-306 (1997); M. Burg, Am. J. Physiol. 268, F983-F996
(1995), each incorporated herein by reference. Although the
inventor does not wish to be bound to any particular theory of the
invention, it appears that these organic osmolytes are useful in
controlling extracellular volume on the airway/pulmonary surface.
Organic osmolytes useful as active compounds in the present
invention include but are not limited to three major classes of
compounds: polyols (polyhydric alcohols), methylamines, and amino
acids. The polyol organic osmolytes considered useful in the
practice of this invention include, but are not limited to,
inositol, myo-inositol, and sorbitol. The methylamine organic
osmolytes useful in the practice of the invention include, but are
not limited to, choline, betaine, carnitine (L-, D- and DL forms),
phosphorylcholine, lyso-phosphorylcholine,
glycerophosphorylcholine, creatine, and creatine phosphate. The
amino acid organic osmolytes of the invention include, but are not
limited to, the D- and L-forms of glycine, alanine, glutamine,
glutamate, aspartate, proline and taurine. Additional osmolytes
useful in the practice of the invention include tihulose and
sarcosine. Mammalian organic osmolytes are preferred, with human
organic osmolytes being most preferred. However, certain organic
osmolytes are of bacterial, yeast, and marine animal origin, and
these compounds are also useful active compounds within the scope
of the present invention.
Under certain circumstances, an osmolyte precursor may be
administered to the subject; accordingly, these compounds are also
useful in the practice of the invention. The term "osmolyte
precursor" as used herein refers to a compound which is converted
into an osmolyte by a metabolic step, either catabolic or anabolic.
The osmolyte precursors of this invention include, but are not
limited to, glucose, glucose polymers, glycerol, choline,
phosphatidylcholine, lyso-phosphatidylcholine and inorganic
phosphates, which are precursors of polyols and methylamines.
Precursors of amino acid osmolytes within the scope of this
invention include proteins, peptides, and polyamino acids, which
are hydrolyzed to yield osmolyte amino acids, and metabolic
precursors which can be converted into osmolyte amino acids by a
metabolic step such as transamination. For example, a precursor of
the amino acid glutamine is poly-L-glutamine, and a precursor of
glutamate is poly-L-glutamic acid.
Also intended within the scope of this invention are chemically
modified osmolytes or osmolyte precursors. Such chemical
modifications involve linking to the osmolyte (or precursor) an
additional chemical group which alters or enhances the effect of
the osmolyte or osmolyte precursor (e.g., inhibits degradation of
the osmolyte molecule). Such chemical modifications have been
utilized with drugs or prodrugs and are known in the art. (See, for
example, U.S. Pat. Nos. 4,479,932 and 4,540,564; Shek, E. et al.,
J. Med. Chem. 19:113-117 (1976); Bodor, N. et al. J. Pharm. Sci.
67:1045-1050 (1978); Bodor, N. et al. J. Med. Chem. 26:313-318
(1983); Bodor, N. et al., J. Pharm. Sci. 75:29-35 (1986), each
incorporated herein by reference.
In general, osmotically active compounds of the present invention
(both ionic and non-ionic) that do not promote, or in fact deter or
retard bacterial growth are preferred.
The compounds of formula (I), formula II, or formula III described
herein and osmotically active compounds disclosed herein may be
administered in any order and/or concurrently to mucosal surfaces
such as the eye, the nose, and airway surfaces including the nasal
passages, sinuses and lungs of a subject by any suitable means
known in the art, such as by nose drops, mists, aerosols,
continuous overnight nasal cannulation, etc. In one embodiment of
the invention, the compounds of formula (I), formula II, or formula
III and osmotically active compounds of the present invention are
administered concurrently by transbronchoscopic lavage. In a
preferred embodiment of the invention, the compounds of formula
(I), formula II, or formula III and osmotically active compounds of
the present invention are deposited on lung airway surfaces by
administering by inhalation an respirable aerosol respirable
particles comprised of the compounds of formula (I), formula II, or
formula III and the osmotically active compounds, in which the
compounds of formula (I), formula II, or formula III can precede or
follow the independent delivery of an osmotically active compound
within a sufficiently short time for their effects to be additive.
The respirable particles may be liquid or solid. Numerous inhalers
for administering aerosol particles to the lungs of a subject are
known. In another preferred embodiment of the invention, the
compounds of formula (I), formula II, or formula III and
osmotically active compounds can be given concurrently as defined
herein.
The compounds of formula (I), formula II, or formula III and
osmotically active compounds of the present invention are
administered sequentially (in any order) or concurrently to the
subject in need thereof. As used herein, the term "concurrently"
means sufficiently close in time to produce a combined effect (that
is, concurrently may be simultaneously, or it may be two or more
events occurring within a short time period before or after each
other). Concurrently also embraces the delivery of the compounds of
formula (I), formula II, or formula III and osmolytes as a mixture
or solution of the two components as well as when delivered from
two different nebulizers. An example of that would be the delivery
of compound I in one nebulizer and hypertonic saline in a second
nebulizer connected by a T-piece. When administered with other
active agents, the active compounds of the present invention may
function as a vehicle or carrier for the other active agent, or may
simply be administered concurrently with the other active agent.
The active compound of the present invention may be used as a dry
or liquid vehicle for administering other active ingredients to
airway surfaces. Such other active agents may be administered for
treating the disease or disorder for which they are intended, in
their conventional manner and dosages, in combination with the
active compounds of the present invention, which may be thought of
as serving as a vehicle or carrier for the other active agent. Any
such other active ingredient may be employed, particularly where
hydration of the airway surfaces (i.e., the activity of the
osmotically active compounds of the present invention) facilitates
the activity of the other active ingredient (e.g., by facilitating
or enhancing uptake of the active ingredient, by contributing to
the mechanism of action of the other active ingredient, or by any
other mechanisms). In a preferred embodiment of the invention, when
the active compound of the present invention is administered
concurrently with another active agent, the active compound of the
present invention has an additive effect in relation to the other
active agent; that is, the desired effect of the other active agent
is enhanced by the concurrent administration of the active
compounds of the present invention.
The compounds of formula (I), formula II, or formula III of the
present invention are also useful for treating airborne infections.
Examples of airborne infections include, for example, RSV. The
compounds of formula (I), formula II, or formula III of the present
invention are also useful for treating an anthrax infection. The
present invention relates to the use of the compounds of formula
(I), formula II, or formula III of the present invention for
prophylactic, post-exposure prophylactic, preventive or therapeutic
treatment against diseases or conditions caused by pathogens. In a
preferred embodiment, the present invention relates to the use of
the compounds of formula (I), formula II, or formula III for
prophylactic, post-exposure prophylactic, preventive or therapeutic
treatment against diseases or conditions caused by pathogens which
may be used in bioterrorism.
In recent years, a variety of research programs and biodefense
measures have been put into place to deal with concerns about the
use of biological agents in acts of terrorism. These measures are
intended to address concerns regarding bioterrorism or the use of
microorganisms or biological toxins to kill people, spread fear,
and disrupt society. For example, the National Institute of Allergy
and Infectious Diseases (NIAID) has developed a Strategic Plan for
Biodefense Research which outlines plans for addressing research
needs in the broad area of bioterrorism and emerging and reemerging
infectious diseases. According to the plan, the deliberate exposure
of the civilian population of the United States to Bacillus
anthracis spores revealed a gap in the nation's overall
preparedness against bioterrorism. Moreover, the report details
that these attacks uncovered an unmet need for tests to rapidly
diagnose, vaccines and immunotherapies to prevent, and drugs and
biologics to cure disease caused by agents of bioterrorism.
Much of the focus of the various research efforts has been directed
to studying the biology of the pathogens identified as potentially
dangerous as bioterrorism agents, studying the host response
against such agents, developing vaccines against infectious
diseases, evaluating the therapeutics currently available and under
investigation against such agents, and developing diagnostics to
identify signs and symptoms of threatening agents. Such efforts are
laudable but, given the large number of pathogens which have been
identified as potentially available for bioterrorism, these efforts
have not yet been able to provide satisfactory responses for all
possible bioterrorism threats. Additionally, many of the pathogens
identified as potentially dangerous as agents of bioterrorism do
not provide adequate economic incentives for the development of
therapeutic or preventive measures by industry. Moreover, even if
preventive measures such as vaccines were available for each
pathogen which may be used in bioterrorism, the cost of
administering all such vaccines to the general population is
prohibitive.
Until convenient and effective treatments are available against
every bioterrorism threat, there exists a strong need for
preventative, prophylactic or therapeutic treatments which can
prevent or reduce the risk of infection from pathogenic agents.
The present invention provides such methods of prophylactic
treatment. In one aspect, a prophylactic treatment method is
provided comprising administering a prophylactically effective
amount of the compounds of formula (I), formula II, or formula III
to an individual in need of prophylactic treatment against
infection from one or more airborne pathogens. A particular example
of an airborne pathogen is anthrax.
In another aspect, a prophylactic treatment method is provided for
reducing the risk of infection from an airborne pathogen which can
cause a disease in a human, said method comprising administering an
effective amount of the compounds of formula (I), formula II, or
formula III to the lungs of the human who may be at risk of
infection from the airborne pathogen but is asymptomatic for the
disease, wherein the effective amount of a sodium channel blocker
and osmolye are sufficient to reduce the risk of infection in the
human. A particular example of an airborne pathogen is anthrax.
In another aspect, a post-exposure prophylactic treatment or
therapeutic treatment method is provided for treating infection
from an airborne pathogen comprising administering an effective
amount of the compounds of formula (I), formula II, or formula III
to the lungs of an individual in need of such treatment against
infection from an airborne pathogen. The pathogens which may be
protected against by the prophylactic post exposure, rescue and
therapeutic treatment methods of the invention include any
pathogens which may enter the body through the mouth, nose or nasal
airways, thus proceeding into the lungs. Typically, the pathogens
will be airborne pathogens, either naturally occurring or by
aerosolization. The pathogens may be naturally occurring or may
have been introduced into the environment intentionally after
aerosolization or other method of introducing the pathogens into
the environment. Many pathogens which are not naturally transmitted
in the air have been or may be aerosolized for use in bioterrorism.
The pathogens for which the treatment of the invention may be
useful includes, but is not limited to, category A, B and C
priority pathogens as set forth by the NIAID. These categories
correspond generally to the lists compiled by the Centers for
Disease Control and Prevention (CDC). As set up by the CDC,
Category A agents are those that can be easily disseminated or
transmitted person-to-person, cause high mortality, with potential
for major public health impact. Category B agents are next in
priority and include those that are moderately easy to disseminate
and cause moderate morbidity and low mortality. Category C consists
of emerging pathogens that could be engineered for mass
dissemination in the future because of their availability, ease of
production and dissemination and potential for high morbidity and
mortality. Particular examples of these pathogens are anthrax and
plague. Additional pathogens which may be protected against or the
infection risk therefrom reduced include influenza viruses,
rhinoviruses, adenoviruses and respiratory syncytial viruses, and
the like. A further pathogen which may be protected against is the
coronavirus which is believed to cause severe acute respiratory
syndrome (SARS).
The compounds of the present invention may also be used in
conjunction with a P2Y2 receptor agonist or a pharmaceutically
acceptable salt thereof (also sometimes referred to as an "active
agent" herein). The composition may further comprise a P2Y2
receptor agonist or a pharmaceutically acceptable salt thereof
(also sometimes referred to as an "active agent" herein). The P2Y2
receptor agonist is typically included in an amount effective to
stimulate chloride and water secretion by airway surfaces,
particularly nasal airway surfaces. Suitable P2Y2 receptor agonists
are described in columns 9-10 of U.S. Pat. No. 6,264,975, U.S. Pat.
No. 5,656,256, and U.S. Pat. No. 5,292,498, each of which is
incorporated herein by reference.
Bronchodilators can also be used in combination with compounds of
the present invention. These bronchodilators include, but are not
limited to, .beta.-adrenergic agonists including but not limited to
epinephrine, isoproterenol, fenoterol, albutereol, terbutalin,
pirbuterol, bitolterol, metaproterenol, iosetharine, salmeterol
xinafoate, as well as anticholinergic agents including but not
limited to ipratropium bromide, as well as compounds such as
theophylline and aminophylline. These compounds may be administered
in accordance with known techniques, either prior to or
concurrently with the active compounds described herein.
Another aspect of the present invention is a pharmaceutical
formulation, comprising an active compound as described above in a
pharmaceutically acceptable carrier (e.g., an aqueous carrier
solution). In general, the active compound is included in the
composition in an amount effective to treat mucosal surfaces, such
as inhibiting the reabsorption of water by mucosal surfaces,
including airway and other surfaces.
The active compounds disclosed herein may be administered to
mucosal surfaces by any suitable means, including topically,
orally, rectally, vaginally, ocularly and dermally, etc. For
example, for the treatment of constipation, the active compounds
may be administered orally or rectally to the gastrointestinal
mucosal surface. The active compound may be combined with a
pharmaceutically acceptable carrier in any suitable form, such as
sterile physiological or dilute saline or topical solution, as a
droplet, tablet or the like for oral administration, as a
suppository for rectal or genito-urethral administration, etc.
Excipients may be included in the formulation to enhance the
solubility of the active compounds, as desired.
The active compounds disclosed herein may be administered to the
airway surfaces of a patient by any suitable means, including as a
spray, mist, or droplets of the active compounds in a
pharmaceutically acceptable carrier such as physiological or dilute
saline solutions or distilled water. For example, the active
compounds may be prepared as formulations and administered as
described in U.S. Pat. No. 5,789,391 to Jacobus, the disclosure of
which is incorporated by reference herein in its entirety.
Solid or liquid particulate active agents prepared for practicing
the present invention could, as noted above, include particles of
respirable or non-respirable size; that is, for respirable
particles, particles of a size sufficiently small to pass through
the mouth and larynx upon inhalation and into the bronchi and
alveoli of the lungs, and for non-respirable particles, particles
sufficiently large to be retained in the nasal airway passages
rather than pass through the larynx and into the bronchi and
alveoli of the lungs. In general, particles ranging from about 1 to
5 microns in size (more particularly, less than about 4.7 microns
in size) are respirable. Particles of non-respirable size are
greater than about 5 microns in size, up to the size of visible
droplets. Thus, for nasal administration, a particle size in the
range of 10-500 .mu.m may be used to ensure retention in the nasal
cavity.
In the manufacture of a formulation according to the invention,
active agents or the physiologically acceptable salts or free bases
thereof are typically admixed with, inter alia, an acceptable
carrier. Of course, the carrier must be compatible with any other
ingredients in the formulation and must not be deleterious to the
patient. The carrier must be solid or liquid, or both, and is
preferably formulated with the compound as a unit-dose formulation,
for example, a capsule, that may contain 0.5% to 99% by weight of
the active compound. One or more active compounds may be
incorporated in the formulations of the invention, which
formulations may be prepared by any of the well-known techniques of
pharmacy consisting essentially of admixing the components.
Compositions containing respirable or non-respirable dry particles
of micronized active agent may be prepared by grinding the dry
active agent with a mortar and pestle, and then passing the
micronized composition through a 400 mesh screen to break up or
separate out large agglomerates.
The particulate active agent composition may optionally contain a
dispersant which serves to facilitate the formulation of an
aerosol. A suitable dispersant is lactose, which may be blended
with the active agent in any suitable ratio (e.g., a 1 to 1 ratio
by weight).
Active compounds disclosed herein may be administered to airway
surfaces including the nasal passages, sinuses and lungs of a
subject by a suitable means know in the art, such as by nose drops,
mists, etc. In one embodiment of the invention, the active
compounds of the present invention and administered by
transbronchoscopic lavage. In a preferred embodiment of the
invention, the active compounds of the present invention are
deposited on lung airway surfaces by administering an aerosol
suspension of respirable particles comprised of the active
compound, which the subject inhales. The respirable particles may
be liquid or solid. Numerous inhalers for administering aerosol
particles to the lungs of a subject are known.
Inhalers such as those developed by Inhale Therapeutic Systems,
Palo Alto, Calif. USA, may be employed, including but not limited
to those disclosed in U.S. Pat. Nos. 5,740,794; 5,654,007;
5,458,135; 5,775,320; and 5,785,049, each of which is incorporated
herein by reference. The Applicant specifically intends that the
disclosures of all patent references cited herein be incorporated
by reference herein in their entirety. Inhalers such as those
developed by Dura Pharmaceuticals, Inc., San Diego, Calif., USA,
may also be employed, including but not limited to those disclosed
in U.S. Pat. Nos. 5,622,166; 5,577,497; 5,645,051; and 5,492,112,
each of which is incorporated herein by reference. Additionally,
inhalers such as those developed by Aradigm Corp., Hayward, Calif.,
USA, may be employed, including but not limited to those disclosed
in U.S. Pat. Nos. 5,826,570; 5,813,397; 5,819,726; and 5,655,516,
each of which is incorporated herein by reference. These
apparatuses are particularly suitable as dry particle inhalers.
Aerosols of liquid particles comprising the active compound may be
produced by any suitable means, such as with a pressure-driven
aerosol nebulizer (L C Star) or an ultrasonic nebulizer (Pari
eFlow). See, e.g., U.S. Pat. No. 4,501,729, which is incorporated
herein by reference. Nebulizers are commercially available devices
which transform solutions or suspensions of the active ingredient
into a therapeutic aerosol mist either by means of acceleration of
compressed gas, typically air or oxygen, through a narrow venturi
orifice or by means of ultrasonic agitation. Suitable formulations
for use in nebulizers consist of the active ingredient in a liquid
carrier, the active ingredient comprising up to 40% w/w of the
formulation, but preferably less than 20% w/w. The carrier is
typically water (and most preferably sterile, pyrogen-free water)
or dilute aqueous alcoholic solution. Perfluorocarbon carriers may
also be used. Optional additives include preservatives if the
formulation is not made sterile, for example, methyl
hydroxybenzoate, antioxidants, flavoring agents, volatile oils,
buffering agents and surfactants.
Aerosols of solid particles comprising the active compound may
likewise be produced with any solid particulate medicament aerosol
generator. Aerosol generators for administering solid particulate
medicaments to a subject produce particles which are respirable, as
explained above, and generate a volume of aerosol containing
predetermined metered dose of medicament at a rate suitable for
human administration. One illustrative type of solid particulate
aerosol generator is an insufflator. Suitable formulations for
administration by insufflation include finely comminuted powders
which may be delivered by means of an insufflator or taken into the
nasal cavity in the manner of a snuff. In the insufflator, the
powder (e.g., a metered dose thereof effective to carry out the
treatments described herein) is contained in capsules or
cartridges, typically made of gelatin or plastic, which are either
pierced or opened in situ and the powder delivered by air drawn
through the device upon inhalation or by means of a
manually-operated pump. The powder employed in the insufflator
consists either solely of the active ingredient or of powder blend
comprising the active ingredient, a suitable powder diluent, such
as lactose, and an optional surfactant. The active ingredient
typically comprises of 0.1 to 100% w/w of the formulation. A second
type of illustrative aerosol generator comprises a metered dose
inhaler. Metered dose inhalers are pressurized aerosol dispensers,
typically containing a suspension or solution formulation of active
ingredient in a liquified propellant. During use, these devices
discharge the formulation through a valve adapted to deliver a
metered volume, typically from 10 to 150 .mu.l, to produce a fine
particle spray containing the active ingredient. Suitable
propellants include certain chlorofluorocarbon compounds, for
example, dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane and mixtures thereof. The formulation may
additionally contain one of more co-solvents, for example, ethanol,
surfactants, such as oleic acid or sorbitan trioleate, antioxidants
and suitable flavoring agents.
The aerosol, whether formed from solid or liquid particles, may be
produced by the aerosol generator at a rate of from about 10 to 150
liters per minute, more preferable from 30 to 150 liters per
minute, and most preferably about 60 liters per minute. Aerosols
containing greater amounts of medicament may be administered more
rapidly.
The dosage of the active compounds disclosed herein will vary
depending on the condition being treated and the state of the
subject, but generally may be from about 0.01, 0.03, 0.05, 0.1 to
1, 5, 10 or 20 mg of the pharmaceutic agent, deposited on the
airway surfaces. The daily dose may be divided among one or
multiple unit dose administrations. The goal is to achieve a
concentration of the pharmaceutic agents on lung airway surfaces of
between 10.sup.-9-10.sup.4 M.
In another embodiment, they are administered by administering an
aerosol suspension of respirable or non-respirable particles
(preferably non-respirable particles) comprised of active compound,
which the subject inhales through the nose. The respirable or
non-respirable particles may be liquid or solid. The quantity of
active agent included may be an amount of sufficient to achieve
dissolved concentrations of active agent on the airway surfaces of
the subject of from about 10.sup.-9, 10.sup.-8, or 10.sup.-7 to
about 10.sup.-3, 10.sup.-2, 10.sup.-1 moles/liter, and more
preferably from about 10.sup.-9 to about 10.sup.-4 moles/liter.
The dosage of active compound will vary depending on the condition
being treated and the state of the subject, but generally may be an
amount sufficient to achieve dissolved concentrations of active
compound on the nasal airway surfaces of the subject from about
10.sup.-9, 10.sup.-8, 10.sup.-7 to about 10.sup.-3, 10.sup.-2, or
10.sup.-1 moles/liter, and more preferably from about 10.sup.-7 to
about 10.sup.-4 moles/liter. Depending upon the solubility of the
particular formulation of active compound administered, the daily
dose may be divided among one or several unit dose administrations.
The daily dose by weight may range from about 0.01, 0.03, 0.1, 0.5
or 1.0 to 10 or 20 milligrams of active agent particles for a human
subject, depending upon the age and condition of the subject. A
currently preferred unit dose is about 0.5 milligrams of active
agent given at a regimen of 2-10 administrations per day. The
dosage may be provided as a prepackaged unit by any suitable means
(e.g., encapsulating a gelatin capsule).
In one embodiment of the invention, the particulate active agent
composition may contain both a free base of active agent and a
pharmaceutically acceptable salt to provide both early release and
sustained release of active agent for dissolution into the mucus
secretions of the nose. Such a composition serves to provide both
early relief to the patient, and sustained relief over time.
Sustained relief, by decreasing the number of daily administrations
required, is expected to increase patient compliance with the
course of active agent treatments.
Pharmaceutical formulations suitable for airway administration
include formulations of solutions, emulsions, suspensions and
extracts. See generally, J. Nairn, Solutions, Emulsions,
Suspensions and Extracts, in Remington: The Science and Practice of
Pharmacy, chap. 86 (19.sup.th ed. 1995), incorporated herein by
reference. Pharmaceutical formulations suitable for nasal
administration may be prepared as described in U.S. Pat. Nos.
4,389,393 to Schor; 5,707,644 to Illum; 4,294,829 to Suzuki; and
4,835,142 to Suzuki, the disclosures of which are incorporated by
reference herein in their entirety.
Mists or aerosols of liquid particles comprising the active
compound may be produced by any suitable means, such as by a simple
nasal spray with the active agent in an aqueous pharmaceutically
acceptable carrier, such as a sterile saline solution or sterile
water. Administration may be with a pressure-driven aerosol
nebulizer or an ultrasonic nebulizer. See e.g. U.S. Pat. Nos.
4,501,729 and 5,656,256, both of which are incorporated herein by
reference. Suitable formulations for use in a nasal droplet or
spray bottle or in nebulizers consist of the active ingredient in a
liquid carrier, the active ingredient comprising up to 40% w/w of
the formulation, but preferably less than 20% w/w. Typically the
carrier is water (and most preferably sterile, pyrogen-free water)
or dilute aqueous alcoholic solution, preferably made in a 0.12% to
0.8% solution of sodium chloride. Optional additives include
preservatives if the formulation is not made sterile, for example,
methyl hydroxybenzoate, antioxidants, flavoring agents, volatile
oils, buffering agents, osmotically active agents (e.g. mannitol,
xylitol, erythritol) and surfactants.
Compositions containing respirable or non-respirable dry particles
of micronized active agent may be prepared by grinding the dry
active agent with a mortar and pestle, and then passing the
micronized composition through a 400 mesh screen to break up or
separate out large agglomerates.
The particulate composition may optionally contain a dispersant
which serves to facilitate the formation of an aerosol. A suitable
dispersant is lactose, which may be blended with the active agent
in any suitable ratio (e.g., a 1 to 1 ratio by weight).
The compounds of formula I-III may be synthesized according to
procedures known in the art. A representative synthetic procedure
is shown in the scheme below:
##STR00042## These procedures are described in, for example, E. J.
Cragoe, "The Synthesis of Amiloride and Its Analogs" (Chapter 3) in
Amiloride and Its Analogs, pp. 25-36, incorporated herein by
reference. Other methods of preparing the compounds are described
in, for example, U.S. Pat. No. 3,313,813, incorporated herein by
reference. See in particular Methods A, B, C, and D described in
U.S. Pat. No. 3,313,813. Additional methods of preparing
intermediates used in the preparation of compounds of the instant
invention are disclosed in U.S. Pat. No. 7,064,129, U.S. Pat. No.
6,858,615, U.S. Pat. No. 6,903,105, WO 2004/073629, WO 2007/146869,
and WO 2007/018640, each of which is expressly incorporated by
reference.
Several assays may be used to characterize the compounds of the
present invention. Representative assays are discussed below.
In Vitro Measure of Sodium Channel Blocking Activity and
Reversibility
One assay used to assess mechanism of action and/or potency of the
compounds of the present invention involves the determination of
lumenal drug inhibition of airway epithelial sodium currents
measured under short circuit current (I.sub.SC) using airway
epithelial monolayers mounted in Ussing chambers. Cells obtained
from freshly excised human, dog, sheep or rodent airways are seeded
onto porous 0.4 micron Snapwell.TM. Inserts (CoStar), cultured at
air-liquid interface (ALI) conditions in hormonally defined media,
and assayed for sodium transport activity (I.sub.SC) while bathed
in Krebs Bicarbonate Ringer (KBR) in Using chambers. All test drug
additions are to the lumenal bath with half-log dose addition
protocols (from 1.times.10.sup.-11 M to 3.times.10.sup.-5 M), and
the cumulative change in I.sub.SC (inhibition) recorded. All drugs
are prepared in dimethyl sulfoxide as stock solutions at a
concentration of 1.times.10.sup.-2 M and stored at -20.degree. C.
Eight preparations are typically run in parallel; two preparations
per run incorporate amiloride and/or benzamil as positive controls.
After the maximal concentration (5.times.10.sup.-5 M) is
administered, the lumenal bath is exchanged three times with fresh
drug-free KBR solution, and the resultant I.sub.SC measured after
each wash for approximately 5 minutes in duration. Reversibility is
defined as the percent return to the baseline value for sodium
current after the third wash. All data from the voltage clamps are
collected via a computer interface and analyzed off-line.
Dose-effect relationships for all compounds are considered and
analyzed by the Prism 3.0 program. IC.sub.50 values, maximal
effective concentrations, and reversibility are calculated and
compared to amiloride and benzamil as positive controls. The
potency of the sodium channel blocking activity of representative
compounds relative to amiloride in freshly excised cell from human
airways is shown in Table 1.
TABLE-US-00001 TABLE 1 Potency of sodium channel blocking activity
of compounds compared to amiloride. Potency of Sodium Channel
Blockade Compound Number Relative to Amiloride 26 30 34 63 72 164
76 347
The potency of the sodium channel blocking activity of
representative compounds in freshly excised cell from dog airways
is shown in Table 2.
TABLE-US-00002 TABLE 2 Compound IC.sub.50 Number nM Amiloride 781
79 23.5 34 18.8 26 25.4 72 6.3 75 4.3 91 6.1 94 9.6 107 8.1 105
23.5 116 2.7 118 3.2
Pharmacological Assays of Absorption
(1) Apical Disappearance Assay
Bronchial cells (dog, human, sheep, or rodent cells) are seeded at
a density of 0.25.times.10.sup.6/cm.sup.2 on a porous Transwell-Col
collagen-coated membrane with a growth area of 1.13 cm.sup.2 grown
at an air-liquid interface in hormonally defined media that
promotes a polarized epithelium. From 12 to 20 days after
development of an air-liquid interface (ALI) the cultures are
expected to be >90% ciliated, and mucins will accumulate on the
cells. To ensure the integrity of primary airway epithelial cell
preparations, the transepithelial resistance (R.sub.t) and
transepithelial potential differences (PD), which are indicators of
the integrity of polarized nature of the culture, are measured.
Human cell systems are preferred for studies of rates of absorption
from apical surfaces. The disappearance assay is conducted under
conditions that mimic the "thin" films in vivo (.about.25 .mu.l)
and is initiated by adding experimental sodium channel blockers or
positive controls (amiloride, benzamil, phenamil) to the apical
surface at an initial concentration of 10 .mu.M. A series of
samples (5 .mu.l volume per sample) is collected at various time
points, including 0, 5, 20, 40, 90 and 240 minutes. Concentrations
are determined by measuring intrinsic fluorescence of each sodium
channel blocker using a Fluorocount Microplate Fluorometer or HPLC.
Quantitative analysis employs a standard curve generated from
authentic reference standard materials of known concentration and
purity. Data analysis of the rate of disappearance is performed
using nonlinear regression, one phase exponential decay (Prism V
3.0).
2. Confocal Microscopy Assay of Amiloride Congener Uptake
Virtually all amiloride-like molecules fluoresce in the ultraviolet
range. This property of these molecules may be used to directly
measure cellular update using x-z confocal microscopy. Equimolar
concentrations of experimental compounds and positive controls
including amiloride and compounds that demonstrate rapid uptake
into the cellular compartment (benzamil and phenamil) are placed on
the apical surface of airway cultures on the stage of the confocal
microscope. Serial x-z images are obtained with time and the
magnitude of fluorescence accumulating in the cellular compartment
is quantitated and plotted as a change in fluorescence versus
time.
3. In vitro Assays of Compound Metabolism
Airway epithelial cells have the capacity to metabolize drugs
during the process of transepithelial absorption. Further, although
less likely, it is possible that drugs can be metabolized on airway
epithelial surfaces by specific ectoenzyme activities. Perhaps more
likely as an ecto-surface event, compounds may be metabolized by
the infected secretions that occupy the airway lumens of patients
with lung disease, e.g. cystic fibrosis. Thus, a series of assays
is performed to characterize the compound metabolism that results
from the interaction of test compounds with human airway epithelia
and/or human airway epithelial lumenal products.
In the first series of assays, the interaction of test compounds in
KBR as an "ASL" stimulant are applied to the apical surface of
human airway epithelial cells grown in the T-Col insert system. For
most compounds, metabolism (generation of new species) is tested
for using high performance liquid chromatography (HPLC) to resolve
chemical species and the endogenous fluorescence properties of
these compounds to estimate the relative quantities of test
compound and novel metabolites. For a typical assay, a test
solution (25 .mu.l KBR, containing 10 .mu.M test compound) is
placed on the epithelial lumenal surface. Sequential 5 to 10 .mu.l
samples are obtained from the lumenal and serosal compartments for
HPLC analysis of (1) the mass of test compound permeating from the
lumenal to serosal bath and (2) the potential formation of
metabolites from the parent compound. In instances where the
fluorescence properties of the test molecule are not adequate for
such characterizations, radiolabeled compounds are used for these
assays. From the HPLC data, the rate of disappearance and/or
formation of novel metabolite compounds on the lumenal surface and
the appearance of test compound and/or novel metabolite in the
basolateral solution is quantitated. The data relating the
chromatographic mobility of potential novel metabolites with
reference to the parent compound are also quantitated.
To analyze the potential metabolism of test compounds by CF sputum,
a "representative" mixture of expectorated CF sputum obtained from
10 CF patients (under IRB approval) has been collected. The sputum
has been be solubilized in a 1:5 mixture of KBR solution with
vigorous vortexing, following which the mixture was split into a
"neat" sputum aliquot and an aliquot subjected to
ultracentrifugation so that a "supernatant" aliquot was obtained
(neat=cellular; supernatant=liquid phase). Typical studies of
compound metabolism by CF sputum involve the addition of known
masses of test compound to "neat" CF sputum and aliquots of CF
sputum "supernatant" incubated at 37.degree. C., followed by
sequential sampling of aliquots from each sputum type for
characterization of compound stability/metabolism by HPLC analysis
as described above. As above, analysis of compound disappearance,
rates of formation of novel metabolites, and HPLC mobilities of
novel metabolites are then performed.
4. Pharmacological Effects and Mechanism of Action of the Drug in
Animals
The effect of compounds for enhancing mucociliary clearance (MCC)
can be measured using an in vivo model described by Sabater et al.,
Journal of Applied Physiology, 1999, pp. 2191-2196, incorporated
herein by reference.
Methods
Animal Preparation:
Adult ewes (ranging in weight from 25 to 35 kg) were restrained in
an upright position in a specialized body harness adapted to a
modified shopping cart. The animals' heads were immobilized and
local anesthesia of the nasal passage was induced with 2%
lidocaine. The animals were then nasally intubated with a 7.5 mm
internal diameter endotracheal tube (ETT). The cuff of the ETT was
placed just below the vocal cords and its position was verified
with a flexible bronchoscope. After intubation the animals were
allowed to equilibrate for approximately 20 minutes prior to
initiating measurements of mucociliary clearance.
Administration of Radio-Aerosol:
Aerosols of .sup.99mTc-Human serum albumin (3.1 mg/ml; containing
approximately 20 mCi) were generated using a Raindrop Nebulizer
which produces a droplet with a median aerodynamic diameter of 3.6
.mu.m. The nebulizer was connected to a dosimetry system consisting
of a solenoid valve and a source of compressed air (20 psi). The
output of the nebulizer was directed into a plastic T connector;
one end of which was connected to the endotracheal tube, the other
was connected to a piston respirator. The system was activated for
one second at the onset of the respirator's inspiratory cycle. The
respirator was set at a tidal volume of 500 mL, an inspiratory to
expiratory ratio of 1:1, and at a rate of 20 breaths per minute to
maximize the central airway deposition. The sheep breathed the
radio-labeled aerosol for 5 minutes. A gamma camera was used to
measure the clearance of .sup.99mTc-Human serum albumin from the
airways. The camera was positioned above the animal's back with the
sheep in a natural upright position supported in a cart so that the
field of image was perpendicular to the animal's spinal cord.
External radio-labeled markers were placed on the sheep to ensure
proper alignment under the gamma camera. All images were stored in
a computer integrated with the gamma camera. A region of interest
was traced over the image corresponding to the right lung of the
sheep and the counts were recorded. The counts were corrected for
decay and expressed as percentage of radioactivity present in the
initial baseline image. The left lung was excluded from the
analysis because its outlines are superimposed over the stomach and
counts can be swallowed and enter the stomach as radio-labeled
mucus.
Treatment Protocol (Assessment of Activity at t-Zero):
A baseline deposition image was obtained immediately after
radio-aerosol administration. At time zero, after acquisition of
the baseline image, vehicle control (distilled water), positive
control (amiloride), or experimental compounds were aerosolized
from a 4 ml volume using a Pari LC JetPlus nebulizer to
free-breathing animals. The nebulizer was driven by compressed air
with a flow of 8 liters per minute. The time to deliver the
solution was 10 to 12 minutes. Animals were extubated immediately
following delivery of the total dose in order to prevent false
elevations in counts caused by aspiration of excess radio-tracer
from the ETT. Serial images of the lung were obtained at 15-minute
intervals during the first 2 hours after dosing and hourly for the
next 6 hours after dosing for a total observation period of 8
hours. A washout period of at least 7 days separated dosing
sessions with different experimental agents.
Treatment Protocol (Assessment of Activity at t-4 Hours):
The following variation of the standard protocol was used to assess
the durability of response following a single exposure to vehicle
control (distilled water), positive control compounds (amiloride or
benzamil), or investigational agents. At time zero, vehicle control
(distilled water), positive control (amiloride), or investigational
compounds were aerosolized from a 4 ml volume using a Pari LC
JetPlus nebulizer to free-breathing animals. The nebulizer was
driven by compressed air with a flow of 8 liters per minute. The
time to deliver the solution was 10 to 12 minutes. Animals were
restrained in an upright position in a specialized body harness for
4 hours. At the end of the 4-hour period animals received a single
dose of aerosolized .sup.99mTc-Human serum albumin (3.1 mg/ml;
containing approximately 20 mCi) from a Raindrop Nebulizer. Animals
were extubated immediately following delivery of the total dose of
radio-tracer. A baseline deposition image was obtained immediately
after radio-aerosol administration. Serial images of the lung were
obtained at 15-minute intervals during the first 2 hours after
administration of the radio-tracer (representing hours 4 through 6
after drug administration) and hourly for the next 2 hours after
dosing for a total observation period of 4 hours. A washout period
of at least 7 days separated dosing sessions with different
experimental agents.
Statistics:
Data were analyzed using SYSTAT for Windows, version 5. Data were
analyzed using a two-way repeated ANOVA (to assess overall
effects), followed by a paired t-test to identify differences
between specific pairs. Significance was accepted when P was less
than or equal to 0.05. Slope values (calculated from data collected
during the initial 45 minutes after dosing in the t-zero
assessment) for mean MCC curves were calculated using linear least
square regression to assess differences in the initial rates during
the rapid clearance phase.
EXAMPLES
Having generally described this invention, a further understanding
can be obtained by reference to certain specific examples which are
provided herein for purposes of illustration only and are not
intended to be limiting unless otherwise specified.
Preparation of Sodium Channel Blockers
Materials and methods. All reagents and solvents were purchased
from Aldrich Chemical Corp. and used without further purification.
Proton and carbon NMR spectra were obtained on a Bruker AC 300
spectrometer at 300 MHz and 75 MHz, respectively. Proton spectra
were referenced to tetramethylsilane as an internal standard and
the carbon spectra were referenced to CDCl.sub.3, CD.sub.3OD,
acetone-d.sub.6 or DMSO-d.sub.6 (purchased from Aldrich or
Cambridge Isotope Laboratories, unless otherwise specified).
Melting points were obtained on a MeI-Temp II apparatus and are
uncorrected. ESI Mass spectra were obtained on a Shimadzu LCMS-2010
EV Mass Spectrometer. HLPC analyses were obtained using a Waters
XTerra RP C18 Analytical Column detected at 220 nm (unless
otherwise specified) on a Shimadzu Prominence HPLC system. With a
flow rate of 1.0 mL per minute, the following time program was
utilized:
TABLE-US-00003 Percent A Percent B Time (H.sub.2O with 0.05% TFA)
(CH.sub.3CN with 0.05% TFA) 0:00 90 10 20:00 10 90 30:00 10 90
35:00 90 10
The following definitions for abbreviations will apply unless
otherwise indicated.
TABLE-US-00004 Abbreviation Definition THF tetrahydrofuran Cbz
Benzyloxycarbonyl i.e. --(CO)O-benzyl AUC Area under the curve or
peak EtOAc Ethyl acetate R.sub.f Retardation factor HPLC High
performance liquid chromatography MTBE Methyl tertiary butyl ether
t.sub.R Retention time GC-MS Gas chromatography-mass spectrometry
wt % Percent by weight h hours min minutes MHz megahertz MeOH
methanol TFA Trifluoroacetic acid UV Ultraviolet
##STR00043## ##STR00044##
Preparation of 4-Bromonaphthol (13)
To a solution of naphthol (12, 5.0 g, 35 mmol) in CH.sub.3CN (125
mL) at 0.degree. C. was added N-bromosuccinimide (7.9 g, 45 mmole)
in several portions. The reaction mixture was warmed to room
temperature, stirred for 1 h, and concentrated. The residue was
dissolved in EtOAc (500 mL) and the solution was washed with water
(300 mL) and brine (300 mL). The organic layer was dried over
MgSO.sub.4, filtered, and concentrated. The residue was purified by
column chromatography (silica gel, 4:1 hexanes/EtOAc) to afford
4-bromonaphthol (13, 5.0 g, 64%) as a white solid: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.22-8.16 (m, 2H), 7.62-7.26 (m, 3H), 6.71
(d, J=8.0 Hz, 1H), 5.46 (s, 1H).
Preparation of
(4-Bromonaphthalen-1-yloxy)(tert-butyl)dimethylsilane (14)
To a solution of imidazole (2.3 g, 34 mmole) and 4-bromonaphthol
(13, 5.0 g, 22 mmole) in DMF (10 mL) at 0.degree. C. was added
t-butyldimethylsilyl chloride (3.7 g, 24.6 mmole) in several
portions. The mixture was warmed to room temperature and stirred
for 2 h. The reaction mixture was partitioned between Et.sub.2O
(500 mL) and water (300 mL) and the aqueous layer was
back-extracted with Et.sub.2O (300 mL). The combined organic layers
were washed with water (300 mL) and brine (300 mL), dried over
MgSO.sub.4, filtered, and concentrated. The residue was purified by
column chromatography (silica gel, hexanes) to afford
(4-bromonaphthalen-1-yloxy)(tert-butyl)dimethylsilane (14, 6.4 g,
85%) as a white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.21-8.14 (m, 2H), 7.61-7.49 (m, 3H), 6.74 (d, J=8.2 Hz, 1H), 1.10
(s, 9H), 0.28 (s, 6H).
Preparation of tert-Butyl (4-Bromonaphthalen-1-yloxy)dimethylsilane
(15)
n-Butyllithium (1.6 M in hexanes, 6.8 mL) was added dropwise to a
solution of (4-bromonaphthalen-1-yloxy)(tert-butyl)dimethylsilane
(14, 3.0 g, 9.0 mmole) in anhydrous THF (30 mL) at -78.degree. C.
and the mixture was stirred for 1 h. Iodine (3.4 g, 14 mmole) in
THF (20 mL) was added dropwise at the same temperature and the
reaction mixture was stirred for 2 h. The reaction mixture was
diluted with Et.sub.2O (500 mL), washed with 1:1 saturated
Na.sub.2S.sub.2O.sub.3/NaHCO.sub.3 (2.times.300 mL) and 1:1
H.sub.2O/brine (300 mL), dried over MgSO.sub.4, filtered, and
concentrated. The residue was purified by column chromatography
(silica gel, hexanes) to afford tert-butyl
(4-bromonaphthalen-1-yloxy)dimethylsilane (15, 1.8 g, 52%) as a
white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.15 (d,
J=8.4 Hz, 1H), 8.02 (d, J=8.6 Hz, 1H), 7.89 (d, J=8.1 Hz, 1H),
7.59-7.47 (m, 2H), 6.64 (d, J=8.1 Hz, 1H), 1.09 (s, 9H), 0.28 (s,
6H).
Preparation of 3-Butyn-1-amine (17)
To a solution of 4-pentynoic acid (16, 15 g, 150 mmole), benzyl
alcohol (17 mL, 170 mmole), and 4-methyl morpholine (17 mL, 150
mmole) in anhydrous toluene (80 mL) was added dropwise diphenyl
phosphoryl azide (33 mL, 150 mmole) at room temperature. The
reaction mixture was stirred for 15 min. The reaction temperature
was carefully raised to 60-70.degree. C., during which vigorous
efflorescence was observed. The reaction mixture was stirred at the
same temperature for 2 h and then at 110.degree. C. for 18 h. The
reaction mixture was cooled to room temperature and concentrated to
a thick brown slurry. The residue was dissolved in CH.sub.2Cl.sub.2
(300 mL) and the solution was stirred for an additional 30 min. The
mixture was washed with water (2.times.300 mL) and the combined
aqueous layers were back-extracted with CH.sub.2Cl.sub.2
(2.times.300 mL). The combined organic layers were dried over
MgSO.sub.4, filtered, and concentrated. The residue was purified by
column chromatography (silica gel, 4:1 hexanes/EtOAc) to afford
amine 17 (16 g, 52%) as a light yellow oil: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.37-7.33 (m, 5H), 5.11 (br s, 3H), 3.36 (q,
J=6.4 Hz, 2H), 2.41 (td, J=6.4, 2.4 Hz, 2H), 1.99 (t, J=2.6 Hz,
1H).
Preparation of Benzyl
4-[4-(tert-Butyldimethylsilyloxy)naphthalen-1-yl]-3-butynylcarbamate
(18)
A solution of tert-butyl (4-bromonaphthalen-1-yloxy)dimethylsilane
(15, 3.1 g, 8.2 mmole), amine 17 (3.3 g, 16 mmole), and triethyl
amine (4.5 mL, 33 mmole) in CH.sub.3CN (70 mL) pre-cooled to
-78.degree. C. was degassed with argon. Tri-tert-butyl phosphine
(10% in hexanes, 3.3 g, 1.6 mmole), Pd(PPh.sub.3).sub.4 (940 mg,
0.82 mmole), and CuI (78 mg, 0.41 mmole) were added rapidly in one
portion at the same temperature. The mixture was warmed to
-30.degree. C. and shaken until a homogeneous solution was formed,
then cooled to -78.degree. C., and degassed with argon. The mixture
was warmed to room temperature and stirred for 18 h. Water (10 mL)
was added to the reaction mixture and the mixture was concentrated.
The residue was diluted with EtOAc (500 mL) and the organic layer
was washed with water (300 mL) and brine (300 mL), dried over
Na.sub.2SO.sub.4, filtered, and concentrated. The residue was
purified by column chromatography (silica gel, 4:1 hexanes/EtOAc)
to afford carbamate 18 (2.0 g, 52%) as a light yellow oil: .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 8.24-8.16 (m, 2H), 7.53-7.45 (m,
3H), 7.38-7.30 (m, 5H), 6.77 (d, J=7.9 Hz, 1H), 5.14 (br s, 3H),
3.56-3.49 (m, 2H), 2.76 (t, J=6.5 Hz, 2H), 1.09 (s, 9H), 0.29 (s,
6H).
Preparation of Benzyl
4-[4-(tert-Butyldimethylsilyloxy)naphthalen-1-yl]butylcarbamate
(19)
A solution of carbamate 18 (2.0 g, 4.3 mmole) and 10% Pd/C (300 mg)
in MeOH (60 mL) was subjected to hydrogenation conditions (50 psi)
for 8 h at room temperature. The reaction mixture was filtered
through a plug of diatomaceous earth and the plug was washed with
MeOH (2.times.20 mL). The filtrate was then concentrated in vacuo
to afford crude amine (1.4 g) which was dissolved in 1:1
CH.sub.2Cl.sub.2/NaHCO.sub.3 (saturated solution) (30 mL). Benzyl
chloroformate (0.62 mL) was added dropwise at room temperature and
the reaction mixture was stirred for 1 h. The mixture was
concentrated, the residue was dissolved in EtOAc (500 mL), and the
solution was washed with water (300 mL) and brine (300 mL). The
organic layer was dried over Na.sub.2SO.sub.4, filtered, and
concentrated. The residue was purified by column chromatography
(silica gel, 4:1 hexanes/EtOAc) to afford butylcarbamate 19 (1.5 g,
77%) as a light yellow oil: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 8.24 (m, 1H), 7.91 (d, J=7.4 Hz, 1H), 7.51-7.42 (m, 2H),
7.34-7.28 (m, 5H), 7.12 (d, J=7.6 Hz, 1H), 6.77 (d, J=7.6 Hz, 1H),
5.08 (br s, 2H), 4.75 (br s, 1H), 3.22 (q, J=6.4 Hz, 2H), 2.99 (q,
J=7.4 Hz, 2H), 1.76-1.71 (m, 2H), 1.67-1.56 (m, 2H), 1.09 (s, 9H),
0.27 (s, 6H).
Preparation of Benzyl 4-(4-Hydroxynaphthalen-1-yl)butylcarbamate
(20)
Tetrabutylammonium fluoride (1 M in THF, 1.0 mL) was added to a
solution of benzyl
4-[4-(tert-butyldimethylsilyloxy)naphthalen-1-yl]butylcarbamate
(19, 380 mg, 0.80 mmol) in anhydrous THF (15 ml) at room
temperature. The reaction mixture was stirred for 2 h and
concentrated to dryness. The residue was purified by column
chromatography (silica gel, 3:1 hexanes/EtOAc) to afford
butylcarbamate 20 (287 mg, 99%) as white solid: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.23-8.21 (m, 1H), 7.84-7.81 (m, 1H),
7.42-7.34 (m, 2H), 7.25-7.15 (m, 5H), 6.96 (d, J=7.5 Hz, 1H), 6.62
(d, J=7.5 Hz, 1H), 5.00 (br s, 2H), 4.65 (br s, 1H), 3.13 (q, J=6.6
Hz, 2H), 2.87 (t, J=7.5 Hz, 2H), 1.64-1.47 (m, 4H).
Preparation of Benzyl
4-[4-(2,3-Dihydroxypropoxy)naphthalen-1-yl]butylcarbamate (22)
A solution of benzyl 4-(4-hydroxynaphthalen-1-yl)butylcarbamate
(20, 287 mg, 0.82 mmole), oxiran-2-ylmethanol (21, 0.07 mL, 1.00
mmole) and triethylamine (0.01 mL, 0.05 mmole) in absolute EtOH
(9.28 mL) was subjected to microwave irradiation at 130.degree. C.
for 30 min. The reaction mixture was concentrated in vacuo and the
residue was purified by column chromatography (silical gel, 95:5
CH.sub.2Cl.sub.2/MeOH) to afford butylcarbamate 22 (293 mg, 83%) as
a light yellow thick oil: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.24 (d, J=7.7 Hz, 1H), 7.94 (d, J=8.1 Hz, 1H), 7.52-7.45 (m, 2H),
7.37-7.28 (m, 5H), 7.18 (d, J=7.7 Hz, 1H), 6.75 (d, J=8.0 Hz, 1H),
5.09 (br s, 2H), 4.71 (br s, 1H), 4.29-4.20 (m, 3H), 3.98-3.81 (m,
2H), 3.23 (q, J=6.5 Hz, 2H), 3.00 (t, J=7.5 Hz, 2H), 2.65 (d, J=4.5
Hz, 1H), 2.06 (t, J=5.8 Hz, 1H), 1.83-1.53 (m, 4H).
Preparation of
3-[4-(4-Aminobutyl)naphthalen-1-yloxy]propane-1,2-diol (23)
A solution of benzyl
4-[4-(2,3-dihydroxypropoxy)naphthalen-1-yl]butylcarbamate (22, 340
mg, 0.80 mmole) and 10% Pd/C (50 mg) in MeOH (50 mL) was subjected
to hydrogenation conditions (1 atm) for 2 h at room temperature.
The reaction mixture was filtered through a plug of diatomaceous
earth and the plug was washed with MeOH. The filtrate was then
concentrated in vacuo to afford diol 23 (226 mg, 97%) as a yellow
solid: MS m/z 290 [C.sub.17H.sub.23NO.sub.3+H].sup.+. Diol 23 was
used in the next step without further purification.
Preparation of
2,4-Diamino-5-chloro-N--{N-[4-(4-(2,3-dihydroxypropoxy)naphthalen-1-yl)bu-
tyl]carbamimidoyl}benzamide (24)
To a solution of
3-[4-(4-aminobutyl)naphthalen-1-yloxy]propane-1,2-diol (23, 226 mg,
0.78 mmole) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 455
mg, 1.17 mmole) in EtOH (10 mL) was added diisopropylethylamine
(0.82 mL, 4.69 mmole) at room temperature. The reaction mixture was
heated at 70.degree. C. in a sealed tube for 7 h, then cooled to
room temperature, and concentrated in vacuo. The residue was
purified by column chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford benzamide 24 (140 mg,
36%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
8.31 (d, J=8.2 Hz, 1H), 7.98 (d, J=8.2 Hz, 1H), 7.53-7.40 (m, 2H),
7.23 (d, J=7.9 Hz, 1H), 6.83 (d, J=7.8 Hz, 1H), 4.22-4.10 (m, 3H),
3.84-3.72 (m, 2H), 3.31-3.25 (m, 2H), 3.08-3.03 (m, 2H), 1.89-1.70
(m, 4H).
Preparation of
3,5-Diamino-6-chloro-N--(N-{4-[6-(2,3-dihydropropoxy)naphthalen-2-yl]buty-
l}carbamimidoyl)pyrazine-2-carboxamide Methanesulfonic Acid Salt
(25)
To a solution of
3,5-diamino-6-chloro-N--(N-{-4-[6-(2,3-dihydropropoxy)naphthalen-2-yl]but-
yl}carbamimidoyl)pyrazine-2-carboxamide (24, 119 mg, 0.24 mmole) in
EtOH (5 mL) was added methanesulfonic acid (22.7 mg, 0.24 mmole) at
room temperature. The reaction mixture was stirred for 15 min. The
solution was concentrated and the residue was azeotroped with MeOH.
The residue was dissolved in H.sub.2O (4 mL) and lyophilized to
afford methanesulfonic acid salt 25 (130 mg, 92%) as a yellow
solid: mp 129-132.degree. C.; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.45 (br s. 1H), 9.12 (br s, 1H), 8.86 (br s, 1H), 8.70
(br s. 1H), 8.27 (dd, J=8.0, 1.3 Hz, 1H), 8.02 (d, J=8.2 Hz, 1H),
7.59-7.49 (m, 2H), 7.47 (br s, 2H), 7.26 (d, J=7.8, 1H), 6.87 (d,
J=7.9 Hz, 1H), 5.06 (d, J=4.7 Hz, 1H), 4.71 (t, J=5.4, 1H),
4.16-3.93 (m, 3H), 3.55 (td, J=5.6, 1.5 Hz, 2H), 3.16 (d. J=5.3 Hz,
2H), 3.00 (t, J=7.0 Hz, 2H), 2.29 (s, 3H), 1.76-1.57 (m, 4H);
ESI-MS m/z 524 [C.sub.23H.sub.28ClN.sub.7O.sub.4+H].sup.+.
Preparation of
2,4-Diamino-5-chloro-N--(N-{4-[4-(2,3-dihydroxypropoxy)naphthalen-1-yl]bu-
tyl}carbamimidoyl)benzamide L-(+)-Lactic Acid Salt (26)
To a solution of
2,4-diamino-5-chloro-N--(N-{4-[4-(2,3-dihydroxypropoxy)naphthalen-1-yl]bu-
tyl}carbamimidoyl)benzamide (24, 28 mg, 0.06 mmole) in EtOH (10 mL)
was added L-(+)-lactic acid (5.20 mg, 0.06 mmole) at room
temperature and the reaction mixture was stirred for 15 min. The
solution was concentrated and the residue was azeotroped with MeOH.
The residue was dissolved in H.sub.2O (3 mL) and lyophilized to
afford lactic acid salt 26 (28 mg, 84%) as a yellow solid: mp
115-118.degree. C.; .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.32
(d, J=8.4 Hz, 1H), 8.01 (d, J=8.4 Hz, 1H), 7.53-7.40 (m, 2H), 7.24
(d, J=7.8 Hz, 1H), 6.83 (d, J=7.8 Hz, 1H), 4.21-4.10 (m, 3H),
4.01-3.94 (m, 1H), 3.82-3.73 (m, 2H), 3.35-3.33 (m, 2H), 3.08 (t,
J=7.0, Hz, 1H), 1.89-1.77 (m, 4H), 1.31 (d, J=7.0 Hz, 3H); ESI-MS
m/z 524 [C.sub.23H.sub.28ClN.sub.7O.sub.4+H].sup.+.
##STR00045## ##STR00046##
Preparation of
(6-Bromonaphthalen-2-yloxy)(tert-butyl)dimethylsilane (28)
A solution of 6-bromonaphthalen-2-ol (5.0 g, 22.4 mmol) and
imidazole (2.3 g, 33.6 mmole) in N,N-dimethylformamide (DMF) (5.0
mL) was added t-butyldimethylsilyl chloride (TBDMSCl) (3.7 g, 24.6
mmole) in one portion at 0.degree. C. The mixture was allowed to
warm to room temperature and stirred for 3 h. The reaction mixture
was partitioned between EtOAc (500 mL) and water (300 mL). The
aqueous layer was separated and extracted with EtOAc (2.times.100
mL) and the combined organic extracts were washed with brine (300
mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated. The
residue was purified by column chromatography (silica gel, hexanes)
to afford (6-bromonaphthalen-2-yloxy)(tert-butyl)dimethylsilane
(28, 7.4 g, 98%) as a white solid: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.91 (d, J=1.8 Hz, 1H), 7.64 (d, J=8.7 Hz, 1H),
7.56 (d, J=8.7 Hz, 1H), 7.48 (dd, J=8.7, 1.8 Hz, 1H), 7.15 (d,
J=2.4 Hz, 1H), 7.09 (dd, J=9.0, 2.4 Hz, 1H), 1.01 (s, 9H), 0.24 (s,
6H).
Preparation of Benzyl
4-[6-(tert-Butyldimethylsilyloxy)naphthalen-2-yl]but-3-ynylcarbamate
(29)
A solution of (6-bromonaphthalen-2-yloxy)(tert-butyl)dimethylsilane
(28, 3.4 g, 10.0 mmol), benzyl but-3-ynylcarbamate (17, 2.0 g, 10
mmole), and triethylamine (20 mL) in anhydrous THF (60 mL)
pre-cooled to -78.degree. C. was degassed with argon. The mixture
was warmed to room temperature and
dichlorobis(triphenylphosphine)palladium(II)
(PdCl.sub.2(PPh.sub.3).sub.2 (702 mg, 1 mmole) and CuI (381 mg, 2
mmole) were added rapidly in one portion under argon. The mixture
was heated at 60.degree. C. for 4 h, then at room temperature for
48 h. The reaction mixture was filtered through a plug of
diatomaceous earth and the filtrate was partitioned between EtOAc
(500 mL) and 1 N HCl (200 mL). The aqueous layer was separated and
back-extracted with EtOAc (300 mL). The combined organic extracts
were washed with water (300 mL) and brine (300 mL), dried over
Na.sub.2SO.sub.4, filtered, and concentrated. The residue was
purified by column chromatography (silical gel, 10:1 hexanes/EtOAc)
to afford carbamate 29 (1.24 g, 37%) as a brown thick oil: .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.83 (s, 1H), 7.65 (d, J=9.0 Hz,
1H), 7.60 (d, J=8.7 Hz, 1H), 7.39-7.29 (m, 6H), 7.13 (d, J=2.1 Hz,
1H), 7.07 (dd, J=8.7, 2.4 Hz, 1H), 5.17 (br s, 1H), 5.13 (s, 2H),
3.46 (q, J=6.3 Hz, 2H), 2.67 (t, J=6.3 Hz, 2H), 1.01 (s, 9H), 0.25
(s, 6H).
Preparation of Benzyl
4-(6-Hydroxynaphthalen-2-yl)but-3-ynylcarbamate (30)
To a solution of benzyl
4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]but-3-ynylcarbamate
(29, 578 mg, 1.26 mmol) in anhydrous THF (60 mL) was added dropwise
tetrabutylammonium fluoride (1 M in THF, 1.38 mL) and the mixture
was stirred for 2 h at room temperature. The resulting solution was
concentrated in vacuo and the residue was purified by column
chromatography (silical gel, 95:5 CH.sub.2Cl.sub.2/MeOH) to afford
carbamate 30 (418 mg, 96%) as a pale yellow solid: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.82 (s, 1H), 7.67 (d, J=9.6 Hz, 1H), 7.58
(d, J=8.4 Hz, 1H), 7.37-7.29 (m, 6H), 7.11-7.08 (m, 2H), 5.30 (br
s, 2H), 5.14 (s, 2H), 3.48 (q, J=6.3 Hz, 2H), 2.68 (t, J=6.6 Hz,
2H).
Preparation of Benzyl
4-[6-(2,3-Dihydroxypropoxy)naphthalen-2-yl]but-3-ynylcarbamate
(31)
A solution of 4-(6-hydroxynaphthalen-2-yl)but-3-ynylcarbamate (30,
390 mg, 1.1 mmole), oxiran-2-ylmethanol (21, 0.1 mL, 1.4 mmole),
and triethylamine (0.01 mL, 0.06 mmole) in absolute EtOH (8.8 mL)
was subjected to microwave irradiation at 130.degree. C. for 30
min. The reaction mixture was concentrated in vacuo and the residue
was purified by column chromatography (silical gel, 95:5
CH.sub.2Cl.sub.2/MeOH) to afford carbamate 31 (236 mg, 42%) as a
white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.80 (s,
1H), 7.68 (dd, J=8.7, 3.9 Hz, 2H), 7.38-7.16 (m, 8H), 5.10 (s, 2H),
4.18 (dd, J=9.6, 4.2 Hz, 1H), 4.11-4.01 (m, 2H), 3.76-3.67 (m, 2H),
3.39-3.31 (m, 4H), 2.63 (t, J=6.9 Hz, 2H).
Preparation of
3-[6-(4-Aminobutyl)naphthalen-2-yloxy]propane-1,2-diol (32)
A suspension of benzyl
4-[6-(2,3-dihydroxypropoxy)naphthalen-2-yl]but-3-ynylcarbamate (31,
236 mg, 0.5 mmole) and 10% Pd/C (96 mg) in MeOH (70 mL) was
subjected to hydrogenation conditions (1 atm) for 1 h at room
temperature. The reaction mixture was filtered through a plug of
diatomaceous earth and the plug was washed with MeOH. The filtrate
was then concentrated in vacuo to afford diol 32 (123 mg, 78%) as a
white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.67 (d,
J=8.4 Hz, 2H), 7.56 (s, 1H), 7.30 (dd, J=8.4, 1.5 Hz, 1H), 7.20 (d,
J=2.4 Hz, 1H), 7.14 (dd, J=8.7, 2.4 Hz, 1H), 4.18-3.99 (m, 3H),
3.76-3.65 (m, 2H), 2.75 (dt, J=10.8, 7.2 Hz, 4H), 1.80-1.70 (m,
2H), 1.62-1.52 (m, 2H).
Preparation of
3,5-Diamino-6-chloro-N--(N-{4-[6-(2,3-dihydropropoxy)naphthalen-2-yl]buty-
l}carbamimidoyl)pyrazine-2-carboxamide (33)
To a solution of
3-[6-(4-aminobutyl)naphthalen-2-yloxy]propane-1,2-diol (32, 51 mg,
0.2 mmole) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 103
mg, 0.3 mmole) in EtOH (2 mL) was added diisopropylethylamine (0.2
mL, 1.1 mmole) at room temperature. The reaction mixture was heated
at 70.degree. C. in a sealed tube for 7 h, then cooled to room
temperature, and concentrated in vacuo. The residue was purified by
column chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carboxamide 33 (34 mg,
68%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.67 (d, J=8.7 Hz, 2H), 7.56 (s, 1H), 7.30 (dd, J=8.4, 1.5 Hz, 1H),
7.19 (d, J=2.4 Hz, 1H), 7.13 (dd, J=9.0, 2.4 Hz, 1H), 4.22-3.99 (m,
3H), 3.76-3.65 (m, 2H), 3.25-3.23 (m, 2H), 2.79 (t, J=7.2 Hz, 2H),
1.84-1.65 (m, 4H).
Preparation of
3,5-Diamino-6-chloro-N--(N-{4-[6-(2,3-dihydropropoxy)naphthalen-2-yl]buty-
l}carbamimidoyl)pyrazine-2-carboxamide Methanesulfonic Acid Salt
(34)
To a solution of
3,5-diamino-6-chloro-N--(N-{4-[6-(2,3-dihydropropoxy)naphthalen-2-yl]buty-
l}carbamimidoyl)pyrazine-2-carboxamide (33, 190 mg, 0.4 mmole) in
EtOH (10 mL) was added methanesulfonic acid (72.7 mg, 0.8 mmole) at
room temperature and the reaction mixture was stirred for 15 min.
The solution was concentrated and the residue was azeotroped with
MeOH. The residue was dissolved in 8:2 MeOH/H.sub.2O (10 mL) and
lyophilized to afford methanesulfonic acid salt 34 (185 mg, 81%) as
a yellow solid: mp 146-149.degree. C.; .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.4 (s, 1H), 9.14 (br s, 1H), 8.87 (br s,
1H), 8.71 (br s, 1H), 7.74 (d, J=8.7 Hz, 2H), 7.62 (s, 1H), 7.44
(br s, 2H), 7.34 (dd, J=8.4, 0.9 Hz, 1H), 7.26 (d, J=2.1 Hz, 1H),
7.14 (dd, J=9.0, 2.4 Hz, 1H), 5.76 (s, 1H), 4.09 (dd, J=9.9, 4.2.
Hz, 1H), 3.98-3.81 (m, 2H), 3.48 (d, J=5.4 Hz, 2H), 3.32 (q, J=5.7
Hz, 2H), 2.75 (t, J=7.2 Hz, 2H), 2.34 (s, 6H), 1.72-1.59 (m, 4H);
ESI-MS m/z 502 [C.sub.23H.sub.28ClN.sub.7O.sub.4+H].sup.+.
##STR00047## ##STR00048##
Preparation of Benzyl 4-(4-Hydroxynaphthalen-1-yl)butylcarbamate
(20)
Tetrabutylammoniumfluoride (1.0 M in THF, 1.1 mL) was added to a
solution of benzyl
4-[4-(tert-butyldimethylsilyloxy)naphthalen-1-yl]butylcarbamate
(19, 500 mg, 1.1 mmole) in THF (5 ml) at room temperature. The
reaction mixture was stirred for 2 h. The reaction mixture was
concentrated to dryness. The residue was purified by column
chromatography (silica gel, 3:1 hexanes/EtOAc) to afford carbamate
20 (230 mg, 61%) as a white solid: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.25-8.21 (m, 1H), 7.91 (dd, J=6.6, 1.8 Hz,
1H), 7.52-7.43 (m, 2H), 7.35-7.27 (m, 5H), 7.05 (d, J=7.5 Hz, 1H),
6.72 (d, J=7.5 Hz, 1H), 5.10 (s, 2H), 4.75 (br s, 1H), 3.23 (q,
J=6.6 Hz, 2H), 2.95 (t, J=7.2 Hz, 2H), 1.76-1.56 (m, 4H).
Preparation of tert-Butyl 3-Hydroxypropylcarbamate (67)
To a solution of 3-aminopropanol (66, 5.0 g, 67 mmole) in 1:1
dioxane/2 N NaOH (100 mL) was added di-tert-butyl dicarbonate (17.0
g, 80 mmole) in dioxane (10 mL) at 0.degree. C. The reaction
mixture was warmed to room temperature and stirred for 1 h. The
mixture was first acidified to pH 1 with concentrated HCl and then
neutralized to pH 7 with 2 N NaOH. The mixture was then extracted
with EtOAc (3.times.200 mL). The combined organic layers were dried
over MgSO.sub.4 and concentrated. The residue was purified by
column chromatography (silica gel, 3:1 hexanes/EtOAc) to afford
tert-butyl 3-hydroxypropylcarbamate (67, 11.0 g, 94%) as a light
yellow oil: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 4.80 (br s,
1H), 3.66 (q, J=5.7 Hz, 2H), 3.33 (q, J=6.3 Hz, 2H), 2.97 (br s,
1H), 1.71-1.63 (m, 2H), 1.45 (s, 9H).
Preparation of Compound (68)
Diisopropylazodicarboxylate (120 mg, 0.59 mmole) was added dropwise
to a solution of benzyl 4-(4-hydroxynaphthalen-1-yl)butylcarbamate
(20, 206 mg, 0.59 mmole), tert-butyl 3-hydroxypropylcarbamate (67,
104 mg, 0.59 mmole), and triphenylphosphine (187 mg, 0.71 mmole) in
anhydrous THF (5 mL) at 0.degree. C. The reaction mixture was
warmed to room temperature and stirred for 5 h. The reaction
mixture was concentrated and the residue was purified by column
chromatography (silica gel, 2:1 hexanes/EtOAc) to afford a mixture
of ether 68 and hydrazine byproduct (630 mg) which was used in the
next step without further purification.
Preparation of tert-Butyl
3-[4-(4-Aminobutyl)naphthalene-1-yloxy]propylcarbamate (69)
A suspension of mixture 68 (630 mg) and 10% Pd/C (300 mg) in MeOH
(25 mL) was subject to hydrogenation conditions (1 atm) for 1 h at
room temperature. The reaction mixture was filtered through a plug
of diatomaceous earth and the plug was washed with MeOH. The
filtrate was concentrated in vacuo and the residue was purified by
column chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carbamate 69 (260 mg,
68% over two steps) as a white solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.28 (dd, J=8.4, 1.2 Hz, 1H), 7.98 (d, J=8.1
Hz, 1H), 7.55-7.42 (m, 2H), 7.22 (d, J=7.8 Hz, 1H), 6.79 (d, J=8.1
Hz, 1H), 4.15 (t, J=6.0 Hz, 2H), 3.32-3.30 (m, 2H), 3.05 (t, J=6.9
Hz, 2H), 2.90 (t, J=7.5 Hz, 2H), 2.11-2.03 (m, 2H), 1.79-1.68 (m,
4H), 1.42 (s, 9H).
Preparation of tert-Butyl
3-{4-[4-(3-(3,5-Diamino-6-chloropyrazine-2-carbonyl)guanidine)butyl]napht-
halen-1-yloxy}propylcarbamate (70)
To a solution of tert-butyl
3-[4-(4-aminobutyl)naphthalene-1-yloxy]propylcarbamate (69, 350 mg,
0.94 mmole) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 600
mg, 1.41 mmole) in EtOH (20 mL) was added diisopropylethylamine
(1.6 mL, 5.2 mmole) at room temperature. The reaction mixture was
heated at 70.degree. C. in a sealed tube for 7 h, cooled to room
temperature and concentrated to dryness. The residue was dissolved
in CHCl.sub.3 (300 mL) and washed with saturated NaHCO.sub.3
(2.times.200 mL). The organic layer was dried over MgSO.sub.4,
filtered, and concentrated. The residue was purified by column
chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carbamate 70 (350 mg,
64%) as a light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.27 (dd, J=8.1, 0.9 Hz, 1H), 7.99 (d, J=8.1 Hz, 1H), 7.50
(td, J=6.6, 1.2 Hz, 1H), 7.42 (td, J=6.6, 1.2 Hz, 1H), 7.23 (d,
J=7.8 Hz, 1H), 6.80 (d, J=7.8 Hz, 1H), 4.17 (t, J=6.0 Hz, 2H),
3.66-3.54 (m, 2H), 3.16-3.05 (m, 4H), 2.12-2.03 (m, 2H), 1.84-1.76
(m, 4H), 1.41 (s, 9H).
Preparation of
3,5-Diamino-N--(N-[4-{4-(3-aminopropoxy)naphthalen-1-yl]butyl}carbamimido-
yl)-6-chloropyrazine-2-carboxamide (71)
To a solution of carbamate 70 (350 mg, 0.6 mmole) in
CH.sub.2Cl.sub.2 (35 mL) was added dropwise trifluoroacetic acid
(2.0 mL) at room temperature. The reaction mixture was stirred for
3 h. The reaction mixture was concentrated in vacuo and azeotroped
with MeOH (2.times.100 mL). The residue was dissolved in water and
the solution was neutralized with saturated NaHCO.sub.3 which
resulted in the precipitation of carboxamide 71. Compound 71 was
collected by filtration and purified by column chromatography
(silica gel, 80:18:2 CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford
amine 71 (185 mg, 64%) as an off-white solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.23 (dd, J=8.1, 0.6 Hz, 1H), 7.99 (d, J=8.1
Hz, 1H), 7.50 (td, J=6.9, 1.5 Hz, 1H), 7.41 (td, J=6.9, 1.2 Hz,
1H), 7.22 (d, J=7.8 Hz, 1H), 6.81 (d, J=7.8 Hz, 1H), 4.20 (t, J=6.3
Hz, 2H), 3.25 (t, J=6.3 Hz, 2H), 3.05 (t, J=6.9 Hz, 2H), 2.97 (t,
J=6.9 Hz, 2H), 2.14-2.05 (m, 2H), 1.89-1.69 (m, 4H).
Preparation of
3,5-Diamino-N--(N-{4-[4-(3-aminopropoxy)naphthalen-1-yl]butyl}carbamimido-
yl)-6-chloropyrazine-2-carboxamide Methanesulfonic Acid Salt
(72)
To a solution of carboxamide 71 (120 mg, 0.247 mmole) in EtOH (10
mL) was added methanesulfonic acid (48 mg, 0.495 mmole) at room
temperature and the reaction mixture was stirred for 15 min. The
solvent was removed in vacuo. The residue was dissolved in water
(10 mL) and lyophilized to afford methanesulfonic acid salt 72 (160
mg, 95%) as a yellow solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.44 (s, 1H), 9.15 (br s, 1H), 8.86 (br s, 1H), 8.72 (br
s, 1H), 8.22 (dd, J=8.1, 0.9, Hz, 1H), 8.03 (d, J=8.1 Hz, 1H), 7.80
(br s, 4H), 7.61-7.49 (m, 3H), 7.42 (br s, 2H), 7.28 (d, J=7.8 Hz,
1H), 6.89 (d, J=7.8 Hz, 1H), 4.22 (t, J=5.7 Hz, 2H), 3.35-3.33 (m,
2H), 3.17-2.99 (m, 4H), 2.32 (s, 6H), 2.20-2.10 (m, 2H), 1.68 (br
s, 4H); ESI-MS m/z 485
[C.sub.23H.sub.29ClN.sub.8O.sub.2+H].sup.+.
##STR00049##
Preparation of Compound (74)
To a solution of amine 71 (60 mg, 0.12 mmole) and Goodman's reagent
73 (100 mg, 0.19 mmole) in MeOH (10 mL) was added
diisopropylethylamine (0.2 mL, 1.0 mmole) at room temperature. The
reaction mixture was stirred for 6 h and then concentrated. The
residue was dissolved in CHCl.sub.3 (100 mL) and washed with
saturated NaHCO.sub.3 (2.times.100 mL). The organic layer was dried
over MgSO.sub.4, filtered, and concentrated. The residue was
purified by column chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford 74 (82 mg, 92%) as a
light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.26
(dd, J=8.1, 0.9 Hz, 1H), 7.96 (d. J=8.1 Hz, 1H), 7.48 (td, J=6.6,
1.2 Hz, 1H), 7.41 (td, J=6.9, 1.2 Hz, 1H), 7.19 (d, J=7.8 Hz, 1H),
6.77 (d, J=7.8 Hz, 1H), 4.19 (t, J=5.7 Hz, 2H), 3.65 (t, J=6.6 Hz,
2H), 3.25 (t, J=6.6 Hz, 2H), 3.01 (t, J=7.2 Hz, 2H), 2.22-2.14 (m,
2H), 1.82-1.65 (m, 4H), 1.41 (s, 9H), 1.42 (s, 9H).
Preparation of Compound (75)
To a solution of compound 74 (130 mg, 0.18 mmole) in
CH.sub.2Cl.sub.2 (20 mL) was added dropwise trifluoroacetic acid
(2.5 mL) at room temperature. The reaction mixture was stirred for
6 h and the solvent was removed in vacuo. The residue was dissolved
in water (10 mL) and the solution was basified to pH 10 with 2 N
NaOH which resulted in the precipitation of crude 75. Compound 75
was collected by filtration and purified by column chromatography
(silica gel, 6:3:1 CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford
compound 75 (48 mg, 51%) as a light yellow solid: .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.27 (dd, J=8.1, 0.9 Hz, 1H), 8.01 (d,
J=8.1 Hz, 1H), 7.53 (td, J=6.9, 1.5 Hz, 1H), 7.45 (td, J=6.9, 1.2
Hz, 1H), 7.26 (d, J=7.8 Hz, 1H), 6.85 (d, J=7.8 Hz, 1H), 4.24 (t,
J=6.0 Hz, 2H), 3.51 (t, J=6.9 Hz, 2H), 3.35-3.33 (m, 2H), 3.08 (t,
J=6.9 Hz, 2H), 2.24-2.19 (m, 2H), 1.84-1.80 (m, 4H).
Preparation of Methanesulfonic Acid Salt (76)
To a solution of compound 75 (48 mg, 0.09 mmole) in EtOH (5 mL) was
added CH.sub.3SO.sub.3H (17.5 mg, 0.18 mmole) at room temperature
and the reaction mixture was stirred for 15 min. The solvent was
removed in vacuo. The residue was dissolved in water (5 mL) and
lyophilized to afford methanesulfonic salt 76 (60 mg, 90%) as a
yellow solid: mp 85-87.degree. C.; .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.44 (s, 1H), 9.15 (br s, 1H), 8.86 (br s,
1H), 8.70 (br s, 1H), 8.24 (dd, J=8.1, 1.5 Hz, 1H), 8.03 (d, J=8.4
Hz, 1H), 7.66-7.42 (m, 6H), 7.42 (br s 2H), 7.28 (d, J=6.9 Hz, 2H),
7.10 (s, 1H), 6.90 (t, J=6.0 Hz, 2H), 4.18 (t, J=6.0 Hz, 2H),
3.43-3.33 (m, 4H), 3.01 (t, J=6.9 Hz, 2H), 2.33 (s, 9H), 2.11-2.06
(m, 2H), 1.68 (br s, 4H); ESI-MS m/z 528
[C.sub.24H.sub.31ClN.sub.10O.sub.2+H].sup.+.
##STR00050##
Preparation of
4-[6-(tert-Butyldimethylsilyloxy)naphthalen-2-yl]butan-1-amine
(77)
A suspension of crude 29 (900 mg) and 10% Pd/C (400 mg) in MeOH (50
mL) was subject to hydrogenation conditions (1 atm) for 6 h at room
temperature. The reaction mixture was filtered through a plug of
diatomaceous earth and the plug was washed with MeOH. The filtrate
was concentrated in vacuo and the residue was purified by column
chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford amine 77 (405 mg, 27%
over two steps) as a white solid: .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.62 (t, J=8.9, 2H), 7.52 (br s, 1H), 7.26 (dd, J=8.4, 1.7
Hz, 1H), 7.15 (d, J=2.2 Hz, 1H), 7.04 (dd, J=8.8, 2.5 Hz, 1H),
3.03-2.37 (br s, 1H), 2.74 (t, J=7.5 Hz, 2H), 1.78-1.65 (m, 2H),
1.58-1.47 (m, 2H), 1.01 (s, 9H), 0.23 (s, 6H).
Preparation of
3,5-Diamino-N--(N-{4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]butyl-
}carbamimidoyl)-6-chloropyrazine-2-carboxamide (78)
To a solution of amine 77 (337 mg, 1.02 mmole) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 596
mg, 1.53 mmole) in EtOH (20 mL) was added diisopropylethylamine
(1.06 mL, 6.13 mmole) at room temperature. The reaction mixture was
heated at 70.degree. C. in a sealed tube for 6 h, then cooled to
room temperature, and concentrated to dryness. The residue was
purified by column chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carboxamide 78 (280 mg,
50%) as a light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 7.67 (d, J=9.2 Hz, 1H), 7.63 (d, J=8.6 Hz, 1H), 7.57 (br s,
1H), 7.30 (d, J=8.1 Hz, 1H), 7.14 (br s, 1H), 7.03 (dd, J=9.2, 1.7
Hz, 1H), 3.30 (m, 2H), 2.81 (t, J=7.0 Hz, 2H), 1.95-1.62 (m, 4H),
1.03 (s, 9H), 0.24 (s, 6H).
Preparation of
3,5-Diamino-6-chloro-N-{N-[4-(6-hydroxynaphthalen-2-yl)butyl]carbamimidoy-
l}pyrazine-2-carboxamide (79)
To a solution of carboxamide 78 (24 mg, 0.05 mmol) in absolute
ethanol (5 mL) was added dropwise 1 N HCl (2 mL) at room
temperature and the mixture was stirred for 12 h. The reaction
mixture was neutralized with saturated NaHCO.sub.3 and compound 79
precipitated out. Compound 79 was collected by filtration and
washed with water (2.times.10 mL) and hexanes (2.times.10 mL) to
afford
3,5-diamino-6-chloro-N--{N-[4-(6-hydroxynaphthalen-2-yl)butyl]carbamimido-
yl}pyrazine-2-carboxamide (79, 10 mg, 53%), after air-drying, as a
light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.63-7.53 (m, 3H), 7.26 (dd, J=8.3, 1.5 Hz, 1H), 7.07-6.98 (m, 2H),
3.36-3.30 (m, 2H), 2.80
##STR00051## ##STR00052##
Preparation of
(6-Bromonaphthalen-2-yloxy)(tert-Butyl)dimethylsilane (81);
A solution of 6-bromonaphthalen-2-ol (12.0 g, 53.7 mmol) and
imidazole (6.0 g, 79.5 mmol) in DMF (12.0 mL) was added
t-butyldimethylsilyl chloride (TBDMSCl) (9.0 g, 59.0 mmol) in one
portion at 0.degree. C. The mixture was allowed to warm to room
temperature and stirred for 3 h. The reaction mixture was
partitioned between EtOAc (500 mL) and water (300 mL). The aqueous
layer was separated and extracted with EtOAc (2.times.100 mL) and
the combined organic extracts were washed with brine (300 mL),
dried over Na.sub.2SO.sub.4, filtered, and concentrated. The
residue was purified by column chromatography (silica gel, hexanes)
to afford (6-bromonaphthalen-2-yloxy)(tert-butyl)dimethylsilane
(81, 18.0 g, 98%) as a white solid: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.91 (d, J=1.8 Hz, 1H), 7.64 (d, J=8.7 Hz, 1H),
7.56 (d, J=8.7 Hz, 1H), 7.48 (dd, J=8.7, 1.8 Hz, 1H), 7.15 (d,
J=2.4 Hz, 1H), 7.09 (dd, J=9.0, 2.4 Hz, 1H), 1.01 (s, 9H), 0.24 (s,
6H).
Preparation of Benzyl
4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]but-3-ynylcarbamate
(83)
A solution of (6-bromonaphthalen-2-yloxy)(tert-butyl)dimethylsilane
(81, 16.1 g, 47.7 mmol), benzyl but-3-ynylcarbamate (82, 9.0 g,
47.7 mmol), and triethylamine (95 mL) in anhydrous THF (100 mL) was
cooled to -78.degree. C. and degassed with argon. The mixture was
warmed to room temperature and
dichlorobis(triphenylphosphine)palladium(II) (3.3 g, 4.8 mmol) and
CuI (1.8 g, 9.6 mmol) were added rapidly in one portion under
argon. The mixture was heated at 50.degree. C. for 12 h. The
reaction mixture was filtered through a plug of diatomaceous earth
and the filtrate was concentrated. The residue was purified by
column chromatography (silical gel, 10:1 hexanes/EtOAc) to afford
carbamate 83 (8.5 g, 38%) as a thick brown oil: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.83 (s, 1H), 7.65 (d, J=9.0 Hz, 1H), 7.60
(d, J=8.7 Hz, 1H), 7.39-7.29 (m, 6H), 7.13 (d, J=2.1 Hz, 1H), 7.07
(dd, J=8.7, 2.4 Hz, 1H), 5.17 (br s, 1H), 5.13 (s, 2H), 3.46 (q,
J=6.3 Hz, 2H), 2.67 (t, J=6.3 Hz, 2H), 1.01 (s, 9H), 0.25 (s,
6H).
Preparation of Benzyl
4-(6-hydroxynaphthalen-2-yl)but-3-ynylcarbamate (84)
To a solution of benzyl
4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]but-3-ynylcarbamate
(83, 2.5 g, 5.44 mmol) in anhydrous THF (25 mL) at 0.degree. C. was
added dropwise tetrabutylammonium fluoride (1 M in THF, 6.0 mL) and
the mixture was stirred for 2 h at room temperature. The resulting
solution was concentrated in vacuo and the residue was purified by
column chromatography (silica gel, 95:5 CH.sub.2Cl.sub.2/MeOH) to
afford carbamate 84 (2.0 g, 50%) as a pale yellow solid: .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.82 (s, 1H), 7.67 (d, J=9.6 Hz,
1H), 7.58 (d, J=8.4 Hz, 1H), 7.37-7.29 (m, 6H), 7.11-7.08 (m, 2H),
5.30 (br s, 2H), 5.14 (s, 2H), 3.48 (q, J=6.3 Hz, 2H), 2.68 (t,
J=6.6 Hz, 2H).
Preparation of tert-Butyl 3-hydroxypropylcarbamate (66)
To a solution of 3-aminopropanol (55, 5.0 g, 67 mmol) in dioxane/2
N NaOH (1:1, 100 mL) was added a solution of di-tert-butyl
dicarbonate (17.0 g, 80 mmol) in dioxane (10 mL) at 0.degree. C.
The reaction mixture was warmed to room temperature and stirred for
1 h. The mixture was first acidified to pH 1 with concentrated HCl
and then neutralized to pH 7 with 2 N NaOH. The mixture was then
extracted with EtOAc (3.times.200 mL). The combined organic layers
were dried over MgSO.sub.4 and concentrated. The residue was
purified by column chromatography (silica gel, 3:1 hexanes/EtOAc)
to afford tert-butyl 3-hydroxypropylcarbamate (86, 11.7 g, 99%) as
a light yellow oil: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 4.80
(br s, 1H), 3.66 (q, J=5.7 Hz, 2H), 3.33 (q, J=6.3 Hz, 2H), 2.97
(br s, 1H), 1.71-1.63 (m, 2H), 1.45 (s, 9H).
Preparation of Boc-protected carbamate (87)
Diisopropylazodicarboxylate (557 mg, 2.75 mmol) was added dropwise
to a solution of 4-(6-hydroxynaphthalen-2-yl)but-3-ynylcarbamate
(84, 638 mg, 1.83 mmol), tert-butyl 3-hydroxypropylcarbamate (86,
355 mg, 2.01 mmol), and triphenylphosphine (980 mg, 3.70 mmol) in
anhydrous THF (20 mL) at 0.degree. C. The reaction mixture was
warmed to room temperature and stirred for 12 h. The reaction
mixture was concentrated and the residue was purified by column
chromatography (silica gel, 2:1 hexanes/EtOAc) to afford a mixture
of ether 87 and the hydrazine by-product (4.0 g) which was used in
the next step without further purification.
Preparation of tert-Butyl
3-[6-(4-aminobutyl)naphthalen-2-yloxy]propylcarbamate (88)
A suspension of 87 (4.0 g) and 10% Pd/C (500 mg) in MeOH/EtOAc
(4:1, 350 mL) was subjected to hydrogenation conditions (1 atm) for
6 h at room temperature. The reaction mixture was filtered through
a plug of diatomaceous earth and the plug was washed with MeOH. The
filtrate was concentrated in vacuo and the residue was purified by
column chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carbamate 88 (437 mg,
64% over two steps) as a white solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.70 (dd, J=9.0 Hz, 2H), 7.59 (s, 1H),
7.34-7.31 (m, 1H), 7.21 (d, J=2.4 Hz, 1H), 7.12 (d, J=9.0, 2.4 Hz,
1H), 4.13 (t, J=6.0 Hz, 2H), 3.30 (t, J=6.3 Hz, 2H), 2.91-2.80 (m,
4H), 2.03-1.99 (m, 2H), 1.90-1.60 (m, 4H), 1.46 (s, 9H).
Preparation of tert-Butyl
3-(6-{4-[3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino]butyl}napht-
halen-2-yloxy)propylcarbamate (89)
To a solution of carbamate 88 (500 mg, 1.34 mmol) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 790
mg, 2.01 mmol) in EtOH (30 mL) was added DIPEA (1.75 mL, 9.39 mmol)
at room temperature. The reaction mixture was heated at 70.degree.
C. in a sealed tube for 2 h, then cooled to room temperature, and
concentrated in vacuo. The residue was purified by column
chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carbamate 89 (660 mg,
84%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.65 (d, J=7.5 Hz, 2H), 7.56 (s, 1H), 7.31 (dd, J=8.4, 1.5 Hz, 1H),
7.16 (d, J=2.4 Hz, 1H), 7.08 (dd, J=8.7, 2.4 Hz, 1H), 4.10 (t,
J=6.3 Hz, 2H), 2.81 (t, J=6.6 Hz, 2H), 2.01-1.97 (m, 2H), 1.83-1.60
(m, 4H), 1.43 (s, 9H).
Preparation of
3,5-Diamino-N--(N-{4-[6-(3-aminopropoxy)naphthalen-2-yl]butyl}carbamimido-
yl-6-chloropyrazine-2-carboxamide (90)
To a solution of compound 89 (725 mg, 1.24 mmol) in
CH.sub.2Cl.sub.2 (45 mL) was added dropwise trifluoroacetic acid
(6.0 mL) at room temperature. The reaction mixture was stirred for
4 h and the solvent was removed in vacuo. The residue was dissolved
in water (10 mL) and the solution was basified to pH 7 with
saturated NaHCO.sub.3 which resulted in the precipitation of crude
20. This was filtered and purified by column chromatography (silica
gel, 80:18:2 CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford compound
90 (289 mg, 48%) as a light yellow solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.65 (d, J=7.5 Hz, 2H), 7.56 (s, 1H), 7.31 (dd,
J=8.4, 1.5 Hz, 1H), 7.16 (d, J=2.4 Hz, 1H), 7.08 (dd, J=8.7, 2.4
Hz, 1H), 4.10 (t, J=6.3 Hz, 2H), 2.81 (t, J=6.6 Hz, 2H), 2.01-1.97
(m, 2H), 1.83-1.60 (m, 4H).
Preparation of
3,5-Diamino-N--(N-{4-[6-(3-aminopropoxy)naphthalen-2-yl]butyl}carbamimido-
yl-6-chloropyrazine-2-carboxamide Methanesulphonate Salt (91)
To a solution of
3,5-diamino-N--(N-{4-[6-(3-aminopropoxy)naphthalen-2-yl]butyl}carbamimido-
yl-6-chloropyrazine-2-carboxamide (20, 30 mg, 0.062 mmol) in EtOH
(5 mL) was added methanesulphonic acid (12.5 mg, 0.13 mmol) at room
temperature and the reaction mixture was stirred for 15 min. The
solution was concentrated and the residue was azeotroped with MeOH.
The residue was dissolved in H.sub.2O/MeOH (8:2, 10 mL) and
lyophilized to afford methanesulphonate salt 91 (33 mg, 79%) as a
yellow solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.4 (s,
1H), 9.13 (br s, 1H), 8.85 (br s, 1H), 8.74 (br s, 1H), 7.76-7.71
(m, 5H), 7.62 (s, 1H), 7.50-7.33 (m, 3H), 7.26 (s, 1H), 7.13 (dd,
J=8.8, 2.2 Hz, 1H), 4.15 (t, J=6.0 Hz, 5H), 3.32-3.04 (m, 2H),
3.01-2.97 (m, 2H), 2.77-2.72 (m, 2H), 2.38 (s, 11H), 2.10-2.01 (m,
2H), 1.74-1.59 (m, 4H); ESI-MS m/z 485
[C.sub.23H.sub.29ClN.sub.8O.sub.2+H].sup.+.
##STR00053## ##STR00054##
Preparation of Boc-urea 92
To a solution of amine 90 (300 mg, 0.62 mmol) and Goodman's reagent
(392 mg, 1.00 mmol) in MeOH (60 mL) was added DIPEA (0.45 mL, 2.5
mmol) at room temperature. The reaction mixture was stirred for 7 h
and then concentrated. The residue was dissolved in CHCl.sub.3 (200
mL) and washed with saturated NaHCO.sub.3 (2.times.100 mL). The
organic layer was dried over MgSO.sub.4, filtered, and
concentrated. The residue was purified by column chromatography
(silica gel, 90:9:1 CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford
Boc-urea 92 (280 mg, 62%) as a light yellow solid: .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 7.67 (t, J=7.8 Hz, 2H), 7.57 (s, 1H), 7.31
(d, J=8.1 Hz, 1H), 7.19 (s, 2H), 4.18 (t, J=5.4 Hz, 2H), 3.62 (t,
J=6.0 Hz, 2H), 2.82 (t, J=6.3 Hz, 2H), 2.12 (t, J=5.7 Hz, 2H),
1.85-1.70 (m, 4H), 1.54 (s, 9H), 1.45 (s, 9H).
Preparation of Urea 93
To a solution of Boc-urea 92 (280 mg, 0.39 mmol) in
CH.sub.2Cl.sub.2 (30 mL) was added dropwise trifluoroacetic acid
(6.0 mL) at room temperature. The reaction mixture was stirred for
4 h and the solvent was removed in vacuo. The residue was dissolved
in water (10 mL) and the solution was basified to pH 10 with 2 N
NaOH which resulted in the precipitation of crude 23. This was
filtered and purified by column chromatography (silica gel, 6:3:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford urea 93 (99 mg, 49%) as
a light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.66 (dd, J=4.5, 8.1 Hz, 2H), 7.57 (s, 1H), 7.32 (d, J=8.1 Hz, 1H),
7.20 (s, 1H), 7.10 (dd, J=2.4, 8.7 Hz, 1H), 4.17 (t, J=5.7 Hz, 2H),
3.43 (t, J=6.6 Hz, 2H), 2.82 (t, J=6.3 Hz, 2H), 2.16-2.08 (m, 2H),
1.84-1.70 (m, 4H).
Preparation of Methanesulphonate Salt 94
To a solution of compound 93 (99 mg, 0.19 mmol) in EtOH (6 mL) was
added CH.sub.3SO.sub.3H (36 mg, 0.40 mmol) at room temperature and
the reaction mixture was stirred for 15 min. The solvent was
removed in vacuo. The residue was dissolved in water (5 mL) and
lyophilized to afford methanesulphonate salt 94 (115 mg, 85%) as a
yellow solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.42 (s,
1H), 9.12 (br s, 1H), 8.85 (br s, 1H), 8.68 (br s, 1H), 7.76-6.90
(m, 16H), 4.10 (t, J=5.4 Hz, 2H), 3.31 (d, J=5.4 Hz, 4H), 2.74-2.71
(m, 2H), 2.30 (s, 6H), 2.07-1.96 (m, 2H), 1.71-1.59 (m, 4H); ESI-MS
m/z 527 [C.sub.24H.sub.31ClN.sub.10O.sub.2+H].sup.+.
##STR00055## ##STR00056##
Preparation of Benzyl
4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]butan-1-amine
(95)
A suspension of 83 (8.0 g, 17.41 mmol) and 10% Pd/C (3.6 g) in MeOH
(240 mL) was subjected to hydrogenation conditions (1 atm) for 6 h
at room temperature. The reaction mixture was filtered through a
plug of diatomaceous earth and the plug was washed with MeOH. The
filtrate was concentrated in vacuo and the residue was purified by
column chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford amine 95 (3.2 g, 56%)
as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.64
(d, J=6.3 Hz, 1H), 7.60 (s, 1H), 7.55 (s, 1H), 7.30 (d, J=1.5 Hz,
1H), 7.15 (s, 1H), 7.03 (dd, J=8.9 Hz, 2.3 Hz, 1H), 2.76 (t, J=7.4
Hz, 2H), 2.70 (t, J=7.2 Hz, 2H), 1.79-1.69 (m, 2H), 1.59-1.49 (m,
2H), 1.04 (s, 9H), 0.25 (s, 6H).
Preparation of Benzyl
4-[6-(tert-butyldimethylsilyloxy)naphthalen-2-yl]carbamate (96)
To a solution of amine 95 (3.2 g, 9.7 mmol) in
CH.sub.2Cl.sub.2/saturated aqueous NaHCO.sub.3 (1:1, 135 mL),
benzyl chloroformate (2.1 mL) was added dropwise at room
temperature and the reaction mixture was stirred for 2 h. The
mixture was concentrated, the residue was dissolved in EtOAc (500
mL), and the solution was washed with water (300 mL) and brine (300
mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered,
and concentrated. The residue was purified by column chromatography
(silica gel, 4:1 hexanes/EtOAc) to afford carbamate 96 (4.0 g, 89%)
as a light yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.64 (d, J=6.3 Hz, 1H), 7.60 (s, 1H), 7.55 (s, 1H), 7.37-7.26 (m,
6H), 7.15 (s, 1H), 7.03 (dd, J=8.9 Hz, 2.3 Hz, 1H), 5.10 (s, 1H),
2.77 (t, J=7.4 Hz, 2H), 2.70 (t, J=7.2 Hz, 2H), 1.79-1.69 (m, 2H),
1.59-1.49 (m, 2H), 1.04 (s, 9H), 0.25 (s, 6H).
Preparation of Benzyl 4-[6-(hydroxynaphthalen-2-yl)]carbamate
(97)
To a solution of carbamate 96 (4.0 g, 6.47 mmol) in THF (30 mL) was
added dropwise tetrabutylammonium fluoride (1 M in THF, 7.2 mL, 7.2
mmol) at room temperature. The reaction mixture was stirred for 2 h
and the solvent was removed in vacuo. The residue was purified by
column chromatography (silica gel, 7:3 hexanes/EtOAc) to afford
compound 97 (2.1 g, 70%) as a light yellow solid: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.63 (d, J=8.7 Hz, 1H), 7.56 (d, J=8.4 Hz,
1H), 7.48 (s, 1H), 7.37-7.26 (m, 5H), 7.21 (dd, J=8.5 Hz, 1.3 Hz,
1H), 7.11 (d, J=2.1 Hz, 1H), 7.08 (d, J=2.4 Hz, 1H), 7.05 (d, J=2.4
Hz, 1H), 5.10 (s, 2H), 4.75 (br, 1H), 3.23 (q, J=6.5 Hz, 2H), 2.72
(t, J=7.5 Hz, 2H), 1.75-1.65 (m, 2H), 1.60-1.47 (m, 2H).
Preparation of Ether 98
Diisopropylazodicarboxylate (2.45 g, 12.0 mmol) was added dropwise
to a solution of benzyl 4-(6-hydroxynaphthalen-2-yl)carbamate (97,
2.1 g, 6.0 mmol), tert-butyl 3-hydroxypropylcarbamate (86, 2.1 g,
12.0 mmol), and triphenylphosphine (4.8 g, 18.0 mmol) in anhydrous
THF (63 mL) at 0.degree. C. The reaction mixture was warmed to room
temperature and stirred for 12 h. The reaction mixture was
concentrated and the residue was purified by column chromatography
(silica gel, 7:3 hexanes/EtOAc) to afford a mixture of ether 98 and
the hydrazine by-product (3.0 g) which was used in the next step
without further purification.
Preparation of Amine 29
To a solution of compound 98 (5.5 g, 11.0 mmol) in CH.sub.2Cl.sub.2
(350 mL) was added dropwise trifluoroacetic acid (84 mL) at room
temperature. The reaction mixture was stirred for 2 h and the
solvent was removed in vacuo. The residue was dissolved in
CHCl.sub.3 (300 mL) and washed with saturated aqueous NaHCO.sub.3,
the organic layer was dried over MgSO.sub.4, filtered, concentrated
in vacuo, and purified by column chromatography (silica gel, 90:9:1
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford compound 99 (1.74 g,
71% over two steps) as a light yellow solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.65 (d, J=8.4 Hz, 2H), 7.53 (s, 1H), 7.31-7.26
(m, 6H), 7.19 (s, 1H), 7.08 (dd, J=8.7 Hz, 2.4 Hz, 1H), 5.05 (s,
2H), 4.17 (t, J=6.0 Hz, 2H), 3.15 (t, J=6.9 Hz, 2H), 2.96 (t, J=6.9
Hz, 2H), 2.73 (t, J=7.5 Hz, 2H), 2.09-2.00 (m, 2H), 1.80-1.65 (m,
2H), 1.59-1.49 (m, 2H).
Preparation of Benzyl
4-[6-(3-{(2S,3R)-2,3-dihydroxy-3-[(4R,5R)-5-hydroxy-2-methyl-1,3-dioxan-4-
-yl]propylamino}propoxy)naphthalene-2-yl]carbamate (101)
A solution of carbamate 99 (1.74 g, 4.28 mmol), triol 100 (922 mg,
4.28 mmol), and sodium triacetoxyborohydride (1.43 g, 6.42 mmol) in
CH.sub.2Cl.sub.2 (18 mL) was stirred at room temperature for 8 h.
The reaction mixture was concentrated to dryness and the residue
was purified by column chromatography (silica gel, 86:12.5:1.5
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH) to afford carbamate 101
(508 mg, 20%) as an white gummy solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.65 (d, J=8.4 Hz, 2H), 7.53 (s, 1H), 7.31-7.26
(m, 6H), 7.19 (s, 1H), 7.11 (dd, J=9.0 Hz, 2.4 Hz, 1H), 5.05 (s,
2H), 4.67 (q, J=4.9 Hz, 1H), 4.18 (t, J=6.0 Hz, 2H), 4.07-3.94 (m,
2H), 3.82-3.74 (m, 2H), 3.46 (dd, J=9.3 Hz, 2.1 Hz, 1H), 3.37 (d,
J=10.5 Hz, 1H), 3.15 (t, J=6.9 Hz, 2H), 3.08-2.72 (m, 6H),
2.13-2.04 (m, 2H), 1.75-1.66 (m, 2H), 1.59-1.52 (m, 2H), 1.25 (d,
J=5.1 Hz, 3H).
Preparation of Benzyl
4-{6-[3-(bis{(2S,3R)-2,3-dihydroxy-3-[(4R,5R)-5-hydroxy-2-methyl-1,3-diox-
an-4-yl]propyl}amino)propoxy]naphthalene-2-yl}carbamate (102)
A solution of carbamate 101 (368 mg, 0.62 mmol), triol 100 (675 mg,
3.10 mmol), sodium cyanoborohydride (338 mg, 4.96 mmol), and HOAc
(290 mg, 4.96 mmol) in MeOH (15 mL) was stirred at room temperature
for 7 d. The reaction mixture was concentrated to dryness; the
residue was washed with saturated NaHCO.sub.3, and extracted with
EtOAc (3.times.200 mL). The organic layers was dried over
MgSO.sub.4, filtered, concentrated, and purified by column
chromatography (silica gel, 80:18:2
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH) to afford carbamate 102
(318 mg, 65%) as an white gummy solid: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.67 (d, J=8.4 Hz, 2H), 7.54 (s, 1H), 7.31-7.27
(m, 6H), 7.21 (s, 1H), 7.11 (dd, J=9.0 Hz, 2.4 Hz, 1H), 5.06 (s,
2H), 4.48 (q, J=5.0 Hz, 2H), 4.17 (t, J=5.7 Hz, 2H), 3.98 (dd,
J=10.5 Hz, 5.4 Hz, 2H), 3.92-3.87 (m, 2H), 3.79-3.72 (m, 4H), 3.35
(d, J=2.1 Hz, 2H), 3.23 (t, J=10.5 Hz, 2H), 3.15 (t, J=6.9 Hz, 2H),
2.82-2.84 (m, 2H), 2.77-2.72 (m, 4H), 2.67-2.60 (m, 2H), 2.03-1.99
(m, 2H), 1.76-1.66 (m, 2H), 1.59-1.52 (m, 2H), 1.20 (d, J=5.1 Hz,
6H).
Preparation of
(R,R,1R,1'R,2S,2'S)-3,3'-{3-[6-(4-Aminobutyl)naphthalene-2-yloxy]propylaz-
anediyl}bis{1-[(4R,5R)-5-hydroxy-2-methyl-1,3-dioxan-4-yl]propane-1,2-diol-
}(103)
A suspension of carbamate 102 (318 mg) and 10% Pd/C (300 mg) in
MeOH (15 mL) was subjected to hydrogenation conditions (1 atm) for
2 h at room temperature. The reaction mixture was filtered through
a plug of diatomaceous earth and the plug was washed with MeOH. The
filtrate was concentrated in vacuo and the residue was purified by
column chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford amine 103 (212 mg, 80%)
as a white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.68
(d, J=7.2 Hz, 2H), 7.56 (s, 1H), 7.30 (d, J=8.1 Hz, 1H), 7.21 (s,
1H), 7.11 (d, J=8.7 Hz, 1H), 4.50-4.49 (m, 2H), 4.17-4.15 (m, 2H),
4.02-3.96 (m, 2H), 3.88 (br s, 2H), 3.79-3.78 (m, 4H), 3.36-3.35
(m, 2H), 7.22 (d, J=10.5 Hz, 2H), 2.80-2.58 (m, 10H), 2.10-1.90 (m,
2H), 1.75-1.73 (m, 2H), 1.65-1.45 (m, 2H), 1.22-1.20 (m, 6H).
Preparation of Carboxamide 104
To a solution of amine 103 (212 mg, 0.33 mmol) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 202
mg, 0.52 mmol) in EtOH (15 mL) was added DIPEA (0.40 mL, 2.28 mmol)
at room temperature. The reaction mixture was heated at 70.degree.
C. in a sealed tube for 2 h, then cooled to room temperature, and
concentrated in vacuo. The residue was purified by column
chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford carboxamide 104 (120
mg, 43%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 7.69-7.65 (m, 2H), 7.57 (s, 1H), 7.30 (d, J=8.1 Hz, 1H),
7.19 (s, 1H), 7.10 (dd, J=9.0 Hz, 2.4 Hz, 1H), 4.50 (q, J=4.8 Hz,
2H), 4.16-4.15 (m, 2H), 4.02-3.96 (m, 2H), 3.90-3.86 (m, 2H),
3.80-3.72 (m, 4H), 3.36-3.20 (m, 6H), 2.83-2.58 (m, 8H), 2.02-1.98
(m, 2H), 1.82-1.79 (m, 2H), 1.71-1.69 (m, 2H), 1.21-1.16 (m,
6H).
Preparation of Carboxamide Lactate Salt 105
To a solution of carboxamide 104 (120 mg, 0.14 mmol) in EtOH (5 mL)
was added lactic acid (27 mg, 0.30 mmol) at room temperature and
the reaction mixture was stirred for 15 min. The solution was
concentrated and the residue was azeotroped with MeOH. The residue
was dissolved in H.sub.2O/MeOH (8:2, 10 mL) and lyophilized to
afford lactate salt 105 (147 mg, >99%) as a yellow solid:
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 7.74 (d, J=8.1 Hz, 2H),
7.63 (s, 1H), 7.34 (d, J=8.4 Hz, 1H), 7.27 (s, 1H), 7.19 (s, 1H),
7.12 (dd, J=9.0 Hz, 2.1 Hz, 1H), 5.14-5.06 (m, Hi), 4.90 (q, J=7.0
Hz, 1H), 4.60 (q, J=5.0 Hz, 2H), 4.23-4.13 (m, 3H), 4.03-3.91 (m,
4H), 3.78-3.77 (m, 2H), 3.70-3.62 (m, 5H), 3.38-3.19 (m, 13H),
2.78-2.58 (m, 8H), 1.97-1.93 (m, 2H), 1.73-1.71 (m, 2H), 1.56-1.62
(m, 2H), 1.47 (d, J=6.9 Hz, 1H), 1.40 (d, J=7.0 Hz, 2H), 1.30 (d,
J=6.9 Hz, 2H), 1.23 (d, J=6.9 Hz, 6H), 1.17 (d, J=5.1 Hz, 6H);
ESI-MS m/z 887 [C.sub.39H.sub.57ClN.sub.8O.sub.12+Na].sup.+.
Alternate Synthesis of
N-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-N'-{4-[4-(3-guanidino-propox-
y)-naphthalen-1-yl]-butyl}-guanidine 75
1. {3-[4-(4-Azido-but-1-enyl)-naphthalen-1-yloxy]-propyl}-carbamic
acid tert-butyl ester
##STR00057##
a. [3-(4-formyl-naphthalen-1-yloxy)-propyl]-carbamic acid
tert-butyl ester
To a solution of 4-hydroxy-naphthalene-1-carbaldehyde (15.2 g, 58.1
mmol) in DMF (50 mL) at r.t. was added N-Boc 3-bromo-propylamine
(15.2 g, 63.9 mmol), followed by potassium carbonate (12 g, 87.2
mmol). The reaction mixture was stirred at room temperature
overnight. Water was added to the reaction mixture and extracted by
CH2Cl2. The combined organic layers were washed with water, brine,
dried over MgSO.sub.4, filtered, and concentrated. The residue was
recrystallized from EtOAc/hexane to afford
[3-(4-formyl-naphthalen-1-yloxy)-propyl]carbamic acid tert-butyl
ester (13.8 g, 72%) as a light yellow solid.
b. {3-[4-(4-Azido-but-1-enyl)-naphthalen-1-yloxy]propyl}-carbamic
acid tert-butyl ester
To a mixture of Wittig reagent
(3-azido-propyl)-triphenyl-phosphonium bromide (15.35 g, 36 mmol)
in THF (150 mL) at -76.degree. C., was added LIHMDS (0.5M in THF
solution, 66 mL, 66 mmol). The mixture was stirred at this
temperature for 1 h.
[3-(4-formyl-naphthalen-1-yloxy)-propyl]-carbamic acid tert-butyl
ester (10 g, 30 mmol) in 20 mL of THF solution was added. The
reaction mixture was stirred for 1 hour. Then warm up to r.t. in 1
h. Water was added to quench the reaction, and extracted with
EtOAc. The organic layer was washed with water, brine, dried over
MgSO.sub.4, filtered, and concentrated. The residue was purified by
column chromatography to afford
{3-[4-(4-azido-but-1-enyl)-naphthalen-1-yloxy]-propyl}-carbamic
acid tert-butyl ester, 11 g, 88% as a solid
c. Wittig reagent (3-azido-propyl)-triphenyl-phosphonium
bromide
##STR00058##
(3-Bromo-propyl)-triphenyl-phosphonium bromide was dissolved in
ethanol/water (1/1). To it sodium azide was added. The reaction
mixture was heated up to reflux overnight. Solvents were removed by
evaporation. The residue was extracted by dry ethanol. Filtered and
evaporated to give crude (3-azido-propyl)-triphenyl-phosphonium
bromide and was used directly for the next step reaction without
further purification.
2.
N-{4-[4-(3-Amino-propoxy)-naphthalen-1-yl]-butyl}-N'-(3,5-diamino-6-chl-
oro-pyrazine-2-carbonyl)-guanidine
##STR00059##
{3-[4-(4-Azido-but-1-enyl)-naphthalen-1-yloxy]-propyl}-carbamic
acid tert-butyl ester 3.5 g was hydrogenated in ethanol with 5%
Pd/C (50% wet) for 2 h. Catalyst was removed, and the filtrate was
concentrated to give 2.94 g of
{3-[4-(4-Amino-butyl)-naphthalen-1-yloxy]-propyl}-carbamic acid
tert-butyl ester.
One gram (2.66 mmol) of free amine was stirred with
1-(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-2-methyl-isothiourea
(1.55 g, 3.99 mmol) in dry ethanol (25 mL). Di-isopropyl-ethylamine
(1.39 mL, 7.98 mmol) of was added and the reaction mixture was
warmed to 45.degree. C. overnight. Ethanol was added and the
reaction filtered. After concentration of the filtrate, the residue
was purified by flash chromatography (0-10% MeOH/CH.sub.2Cl.sub.2)
to give 0.92 g of
[3-(4-{4-[N'-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-guanidino]-butyl}-
-naphthalen-1-yloxy)-propyl]-carbamic acid tert-butyl ester.
[3-(4-{4-[N'-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-guanidino]-butyl}-
-naphthalen-1-yloxy)-propyl]-carbamic acid tert-butyl ester (2.7 g
was stirred with 4M HCl in p-dioxane for 1 hour at room
temperature. Solvents were removed in vacuo.
A small amount of the product was purified by flash chromatography
to give 7 GS-426675
N-{4-[4-(3-Amino-propoxy)-naphthalen-1-yl]-butyl}-N'-(3,5-diamino-6-chlor-
o-pyrazine-2-carbonyl)-guanidine as an HCl salt.
3.
N-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-N'{4-[4-(3-guanidino-propo-
xy)-naphthalen-1-yl]-butyl}-guanidine 75
##STR00060##
N-{4-[4-(3-Amino-propoxy)-naphthalen-1-yl]-butyl}-N'-(3,5-diamino-6-chlor-
o-pyrazine-2-carbonyl)-guanidine HCl salt from reaction was stirred
with Goodman's reagent [(tert-butoxycarbonylamino-trifluoromethane
sulfonylimino-methyl)-carbamic acid tert-butyl ester] in methanol.
Diisopropylethylamine (1.18 mL) was added and the reaction mixture
was stirred at room temperature overnight. Solvents were removed in
vacuo and the residue was purified by silica gel chromatography
(0-10% MeOH(MeOH/NH.sub.4OH=9/1)/CH.sub.2Cl.sub.2) to give 2.7 g of
8, which was dissolve in 30 mL of methanol and treated with 300 mL
of 4M HCl in p-dioxane at room temperature for 4 hour to give 9,
N-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-N'-{4-[4-(3-guanidino-propox-
y)-naphthalen-1-yl]-butyl}-guanidine as a crude product. About 200
mL of the solvents were removed by reduced pressure, then cool to
room temperature, and let the product precipitated out. Filtration
to collect the product, and the product was further refluxed with
dry EtOH, and cool down to room temperature. Filtration to give
2.08 g of 75 as an HCl salt (yellow solid).
##STR00061## ##STR00062##
Preparation of
(1R,2S)-3-{3-[6-(4-Aminobutyl)naphthalene-2-yloxy]propylamino}-1-[(4R,5R)-
-5-hydroxy-2-methyl-1,3-dioxan-4-yl]propane-1,2-diol} (105)
A suspension of 101 (76 mg) and 10% Pd/C (76 mg) in MeOH (5 mL) was
subjected to hydrogenation conditions (1 atm) for 2 h at room
temperature. The reaction mixture was filtered through a plug of
diatomaceous earth and the plug was washed with MeOH. The filtrate
was concentrated in vacuo and the residue was purified by column
chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford amine 105 (46 mg, 80%)
as a white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) .quadrature.
7.65 (d, J=8.7 Hz, 2H), 7.53 (s, 1H), 7.28 (d, J=8.4 Hz, 1H), 7.18
(s, 1H), 7.10 (dd, J=8.7 Hz, 1.8 Hz, 1H), 4.66 (q, J=4.8 Hz, 1H),
4.14 (t, J=6.0 Hz, 2H), 4.06 (q, J=5.3 Hz, 1H), 3.98-3.92 (m, 1H),
3.83-3.74 (m, 2H), 3.45 (dd, J=9.0 Hz, 1.5 Hz, 1H), 3.37 (d, J=10.5
Hz, 1H), 2.93-2.66 (m, 8H), 2.08-2.00 (m, 2H), 1.77-1.67 (m, 2H),
1.58-1.49 (m, 2H), 1.25 (d, J=4.8 Hz, 3H).
Preparation of Guanidine 106
To a solution of amine 105 (46 mg, 0.10 mmol) and methyl
3,5-diamino-6-chloropyrazine-2-carbonylcarbamimidothioate (10, 70
mg, 0.16 mmol) in EtOH (6 mL) was added DIPEA (0.13 mL, 0.7 mmol)
at room temperature. The reaction mixture was heated at 70.degree.
C. in a sealed tube for 2 h, then cooled to room temperature, and
concentrated in vacuo. The residue was purified by column
chromatography (silica gel, 80:18:2
CHCl.sub.3/CH.sub.3OH/NH.sub.4OH) to afford guanidine 106 (16 mg,
24%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD)
.quadrature. 7.66 (d, J=7.5 Hz, 2H), 7.55 (s, 1H), 7.30 (d, J=8.1
Hz, 1H), 7.17 (s, 1H), 7.10 (d, J=8.7 Hz, 1H), 4.67-4.66 (m, 1H),
4.16 (s, 2H), 4.08-3.97 (m, 2H), 3.82-3.77 (m, 2H), 3.64-3.58 (m,
2H), 3.46 (d, J=9.0 Hz, 1H). 3.40 (d, J=3.3 Hz, 1H), 2.05-2.09 (m,
2H), 1.82-1.70 (m, 4H), 1.25 (d, J=4.8 Hz, 3H).
Preparation of Guanidine Lactate Salt 107
To a solution of guanidine 106 (16 mg, 0.024 mmol) in EtOH (5 mL)
was added lactic acid (4.5 mg, 0.048 mmol) at room temperature and
the reaction mixture was stirred for 15 min. The solution was
concentrated and the residue was azeotroped with MeOH. The residue
was dissolved in H.sub.2O/MeOH (8:2, 10 mL) and lyophilized to
afford lactate salt 107 (20 mg, >95%) as a yellow solid: .sup.1H
NMR (300 MHz, CD.sub.3OD) .quadrature. 7.78 (t, J=7.8 Hz, 2H), 7.62
(s, 1H), 7.35 (d, J=8.4 Hz, 1H), 7.27 (s, 1H), 7.13 (dd, J=8.8 Hz,
2.3 Hz, 1H), 7.00 (br s, 1H), 5.10-5.05 (m, 1H), 4.85 (q, J=7.0 Hz,
1H), 4.63 (q, J=5.0 Hz, 1H), 4.21-4.12 (m, 3H), 3.99-3.87 (m, 4H),
3.73 (d, J=5.1 Hz, 1H), 3.68-3.59 (m, 2H), 3.44-3.23 (m, 4H),
3.06-3.00 (m, 4H), 2.90-2.83 (m, 1H), 2.75 (t, J=6.7 Hz, 2H),
2.11-2.07 (m, 2H), 1.71-1.57 (m, 4H), 1.46 (d, J=6.9 Hz, 1H), 1.37
(d, J=7.2 Hz, 3H), 1.28 (d, J=6.6 Hz, 3H), 1.24-1.13 (m, 10H),
0.86-0.82 (m, 1H); ESI-MS m/z 675
[C.sub.31H.sub.43ClN.sub.8O.sub.7+H].sup.+.
Preparation of
N-{4-[4-(3-Amino-propoxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-N'-(-
3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidine (116) and
N-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-N'-{4-[4-(3-guanidino-propox-
y)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-guanidine (118)
1. 4-hydroxy-5,6,7,8-tetrahydro-naphthalene-1-carbaldehyde
(109)
##STR00063##
5,6,7,8-Tetrahydro-naphthalen-1-ol (108, 20 g, 135 mmol) was
stirred in 100 mL of ethanol, and potassium hydroxide (7.57 g, 135
mmol) as an aqueous solution was added. The mixture was stirred for
15 minutes and went clear. Solvents were removed and dried. PEG (MW
380-420, 53 mL) was added, followed by chloroform (32.3 mL, 405
mmol) and toluene (34 mL). An aqueous potassium hydroxide solution
(50% by weight, 54.4 mL) was introduced dropwise with stirring over
15 minutes. The stirring was continued for another 30 minutes. 1M
HCl was added to acidify the reaction mixture and it was extracted
with EtOAc three times. The combined organic layers were washed
with water and brine The combined organic layers were dried over
MgSO.sub.4, filtered, concentrated, and purified by flash
chromatography (0-40% EtOAc/hexane) to give
4-hydroxy-5,6,7,8-tetrahydro-naphthalene-1-carbaldehyde (109, 4.7
g).
2.
[3-(4-Formyl-5,6,7,8-tetrahydro-naphthalen-1-yloxy)-propyl]-carbamic
acid tert-butyl ester (111)
##STR00064##
4-Hydroxy-5,6,7,8-tetrahydro-naphthalene-1-carbaldehyde (109, 4.6
g, 29.1 mmol) (3-Bromopropyl)-carbamic acid tert-butyl ester (110,
4.6 g, 32 mmol), and potassium carbonate (6.03 g, 43.7 mmol) were
stirred in 140 mL of dry DMF over night. The reaction mixture was
poured into water and extracted with dichloromethane. The organic
layer was washed with water and brine., dried with magnesium
sulfate, filtered, concentrated and purified by flash
chromatography (0-30% EtOAc/hexane) to give crude product, which
was recrystallized from EtOAc/Hexane to give 5.8 g of 111,
[3-(4-formyl-5,6,7,8-tetrahydro-naphthalen-1-yloxy)-propyl]-carbamic
acid tert-butyl ester.
3.
{3-[4-(4-Azido-but-1-enyl)-5,6,7,8-tetrahydro-naphthalen-1-yloxy]-propy-
l}-carbamic acid tert-butyl ester (113)
##STR00065##
(3-Azido-propyl)-triphenylphosphonium bromide (112, 11.5 g, 27
mmol) was stirred with 100 mL of dry THF at -76.degree. C. LiHMDS
(0.5 M in toluene, 27 mL) was added and the mixture was stirred for
30 minutes.
[3-(4-Formyl-5,6,7,8-tetrahydro-naphthalen-1-yloxy)-propyl]-carbamic
acid tert-butyl ester (111, 6 g, 18 mmol) in 12 mL dry THF solution
was introduced. The reaction mixture was stirred at this
temperature for another 30 minutes, and slowly warmed to room
temperature. The mixture was poured into water and extracted twice
with ethyl acetate. The combined organic layers were washed with
water and brine, dried over magnesium sulfate, filtered,
concentrated, and purified by flash chromatography (0-25%
EtOAc/Hexane) to give 3.5 g 113
{3-[4-(4-azido-but-1-enyl)-5,6,7,8-tetrahydro-naphthalen-1-yloxy]-propyl}-
-carbamic acid tert-butyl ester.
4.
N-{4-[4-(3-Amino-propoxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-N'-
-(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidine (116)
##STR00066##
{3-[4-(4-Azido-but-1-enyl)-5,6,7,8-tetrahydro-naphthalen-1-yloxy]-propyl}-
-carbamic acid tert-butyl ester 113, 3.5 g was hydrogenated in
ethanol with 5% Pd/C (50% wet) for 2 h. Catalyst was removed, and
the filtrate was concentrated to give 2.94 g of 114,
{3-[4-(4-amino-butyl)-5,6,7,8-tetrahydro-naphthalen-1-yloxy]-propyl}-carb-
amic acid tert-butyl ester.
One gram (2.66 mmol) of free amine 114 was stirred with
1-(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-2-methyl-isothiourea
(1.55 g, 3.99 mmol) in dry ethanol (25 mL). Di-isopropyl-ethylamine
(1.39 mL, 7.98 mmol) of was added and the reaction mixture was
warmed to 45.degree. C. overnight. Ethanol was added and the
reaction filtered. After concentration of the filtrate, the residue
was purified by flash chromatography (0-10% MeOH/CH.sub.2Cl.sub.2)
to give 0.92 g of 115
[3-(4-{4-[N'-(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidino]-butyl}-
-5,6,7,8-tetrahydro-naphthalen-1-yloxy)-propyl]-carbamic acid
tert-butyl ester.
N-{4-[4-(3-Amino-propoxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-N'-(-
3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidine was stirred
with 4M HCl in p-dioxane for 30 minutes at room temperature.
Solvents were removed n vacuo, and the product was purified by
amine column (0-40% MeOH/CH.sub.2Cl.sub.2) to give 116
N-{4-[4-(3-amino-propoxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-N'-(-
3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidine as an HCl
salt.
5.
N-(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-N'-{4-[4-(3-guanidino-prop-
oxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-guanidine GS-429269
(11)
##STR00067##
N-{4-[4-(3-Amino-propoxy)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-N'-(-
3,5-diamino-6-chloro-pyrazine-2-carbonyl)-guanidine HCl salt 116
(0.81 g, 1.37 mmol) was stirred with Goodman's reagent
[(tert-butoxycarbonylamino-trifluoromethane
sulfonylimino-methyl)-carbamic acid tert-butyl ester] in methanol.
Diisopropylethylamine (1.18 mL) was added and the reaction mixture
was stirred at room temperature overnight. Solvents were removed in
vacuo and the residue was purified by silica gel chromatography
(0-10% MeOH/CH.sub.2Cl.sub.2) to give 820 mg of 117, which was
treated with 4M HCl in p-dioxane at room temperature for 1 hour to
give 118,
N-(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-N'-{4-[4-(3-guanidino-propox-
y)-5,6,7,8-tetrahydro-naphthalen-1-yl]-butyl}-guanidine as a crude
product. Purification by flash chromatography (0-40% (MeOH/NH4OH;
3/1)/CH.sub.2Cl.sub.2) followed by further purification on an amine
column (0-30% MeOH/CH.sub.2Cl.sub.2) gave the free base, which was
dissolved in ethanol and a few drops of 1M HCl aq was added. The
clear solution was filtered and lyophilized to give final product
as a yellow solid.
All references cited herein are hereby incorporated in their
entirety as if each reference was individually and specifically
incorporated in its entirety.
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